Compositions and methods relating to breast specific genes and proteins

ABSTRACT

The present invention relates to newly identified nucleic acids and polypeptides present in normal and neoplastic breast cells, including fragments, variants and derivatives of the nucleic acids and polypeptides. The present invention also relates to antibodies to the polypeptides of the invention, as well as agonists and antagonists of the polypeptides of the invention. The invention also relates to compositions comprising the nucleic acids, polypeptides, antibodies, variants, derivatives, agonists and antagonists of the invention and methods for the use of these compositions. These uses include identifying, diagnosing, monitoring, staging, imaging and treating breast cancer and non-cancerous disease states in breast tissue, identifying breast tissue, monitoring and identifying and/or designing agonists and antagonists of polypeptides of the invention. The uses also include gene therapy, production of transgenic animals and cells, and production of engineered breast tissue for treatment and research.

[0001] This application claims the benefit of priority from U.S.Provisional Application Serial No. 60/243,805 filed Oct. 27, 2000, whichis herein incorporated by reference in its entirety.

FIELD OF THE INVENTION

[0002] The present invention relates to newly identified nucleic acidmolecules and polypeptides present in normal and neoplastic breastcells, including fragments, variants and derivatives of the nucleicacids and polypeptides. The present invention also relates to antibodiesto the polypeptides of the invention, as well as agonists andantagonists of the polypeptides of the invention. The invention alsorelates to compositions comprising the nucleic acids, polypeptides,antibodies, variants, derivatives, agonists and antagonists of theinvention and methods for the use of these compositions. These usesinclude identifying, diagnosing, monitoring, staging, imaging andtreating breast cancer and non-cancerous disease states in breasttissue, identifying breast tissue and monitoring and identifying and/ordesigning agonists and antagonists of polypeptides of the invention. Theuses also include gene therapy, production of transgenic animals andcells, and production of engineered breast tissue for treatment andresearch.

BACKGROUND OF THE INVENTION

[0003] Excluding skin cancer, breast cancer, also called mammary tumor,is the most common cancer among women, accounting for a third of thecancers diagnosed in the United States. One in nine women will developbreast cancer in her lifetime and about 192,000 new cases of breastcancer are diagnosed annually with about 42,000 deaths. Bevers, PrimaryPrevention of Breast Cancer, in Breast Cancer, 20-54 (Kelly K Hunt etal., ed., 2001); Kochanek et al., 49 Nat'l.Vital Statistics Reports 1,14 (2001).

[0004] In the treatment of breast cancer, there is considerable emphasison detection and risk assessment because early and accurate staging ofbreast cancer has a significant impact on survival. For example, breastcancer detected at an early stage (stage T0, discussed below) has afive-year survival rate of 92%. Conversely, if the cancer is notdetected until a late stage (i.e., stage T4), the five-year survivalrate is reduced to 13%. AJCC Cancer Staging Handbook pp. 164-65 (IrvinD. Fleming et al. eds., 5^(th) ed. 1998). Some detection techniques,such as mammography and biopsy, involve increased discomfort, expense,and/or radiation, and are only prescribed only to patients with anincreased risk of breast cancer.

[0005] Current methods for predicting or detecting breast cancer riskare not optimal. One method for predicting the relative risk of breastcancer is by examining a patient's risk factors and pursuing aggressivediagnostic and treatment regiments for high risk patients. A patient'srisk of breast cancer has been positively associated with increasingage, nulliparity, family history of breast cancer, personal history ofbreast cancer, early menarche, late menopause, late age of first fullterm pregnancy, prior proliferative breast disease, irradiation of thebreast at an early age and a personal history of malignancy. Lifestylefactors such as fat consumption, alcohol consumption, education, andsocioeconomic status have also been associated with an increasedincidence of breast cancer although a direct cause and effectrelationship has not been established. While these risk factors arestatistically significant, their weak association with breast cancerlimited their usefulness. Most women who develop breast cancer have noneof the risk factors listed above, other than the risk that comes withgrowing older. NIH Publication No. 00-1556 (2000).

[0006] Current screening methods for detecting cancer, such as breastself exam, ultrasound, and mammography have drawbacks that reduce theireffectiveness or prevent their widespread adoption. Breast self exams,while useful, are unreliable for the detection of breast cancer in theinitial stages where the tumor is small and difficult to detect bypalpitation. Ultrasound measurements require skilled operators at anincreased expense. Mammography, while sensitive, is subject to overdiagnosis in the detection of lesions that have questionable malignantpotential. There is also the fear of the radiation used in mammographybecause prior chest radiation is a factor associated with an increaseincidence of breast cancer.

[0007] At this time, there are no adequate methods of breast cancerprevention. The current methods of breast cancer prevention involveprophylactic mastectomy (mastectomy performed before cancer diagnosis)and chemoprevention (chemotherapy before cancer diagnosis) which aredrastic measures that limit their adoption even among women withincreased risk of breast cancer. Bevers, supra.

[0008] A number of genetic markers have been associated with breastcancer. Examples of these markers include carcinoembryonic antigen (CEA)(Mughal et al., 249 JAMA 1881 (1983)) MUC-1 (Frische and Liu, 22 J.Clin. Ligand 320 (2000)), HER-2/neu (Haris et al., 15Proc.Am.Soc.Clin.Oncology. A96 (1996)), uPA, PAI-1, LPA, LPC, RAK andBRCA (Esteva and Fritsche, Serum and Tissue Markers for Breast Cancer,in Breast Cancer, 286-308 (2001)). These markers have problems withlimited sensitivity, low correlation, and false negatives which limittheir use for initial diagnosis. For example, while the BRCA1 genemutation is useful as an indicator of an increased risk for breastcancer, it has limited use in cancer diagnosis because only 6.2% ofbreast cancers are BRCA1 positive. Malone et al., 279 JAMA 922 (1998).See also, Mewman et al., 279 JAMA 915 (1998) (correlation of only 3.3%).

[0009] Breast cancers are diagnosed into the appropriate stagecategories recognizing that different treatments are more effective fordifferent stages of cancer. Stage TX indicates that primary tumor cannotbe assessed (i.e., tumor was removed or breast tissue was removed).Stage T0 is characterized by abnormalities such as hyperplasia but withno evidence of primary tumor. Stage Tis is characterized by carcinoma insitu, intraductal carcinoma, lobular carcinoma in situ, or Paget'sdisease of the nipple with no tumor. Stage T1 is characterized as havinga tumor of 2 cm or less in the greatest dimension. Within stage T1, Tmicindicates microinvasion of 0.1 cm or less, T1a indicates a tumor ofbetween 0.1 to 0.5 cm, T1b indicates a tumor of between 0.5 to 1 cm, andT1c indicates tumors of between 1 cm to 2 cm. Stage T2 is characterizedby tumors from 2 cm to 5 cm in the greatest dimension. Tumors greaterthan 5 cm in size are classified as stage T4. Within stage T4, T4aindicates extension of the tumor to the chess wall, T4b indicates edemaor ulceration of the skin of the breast or satellite skin nodulesconfined to the same breast, T4c indicates a combination of T4a and T4b,and T4d indicates inflammatory carcinoma. AJCC Cancer Staging Handbookpp. 159-70 (Irvin D. Fleming et al. eds., 5^(th) ed. 1998). In additionto standard staging, breast tumors may be classified according to theirestrogen receptor and progesterone receptor protein status. Fisher etal., 7 Breast Cancer Research and Treatment 147 (1986). Additionalpathological status, such as HER2/neu status may also be useful. Thor etal., 90 J.Nat'l.Cancer Inst. 1346 (1998); Paik et al., 90 J.Nat'l.CancerInst. 1361 (1998); Hutchins et al., 17 Proc.Am.Soc.Clin.Oncology A2(1998).; and Simpson et al., 18 J.Clin.Oncology 2059 (2000).

[0010] In addition to the staging of the primary tumor, breast cancermetastases to regional lymph nodes may be staged. Stage NX indicatesthat the lymph nodes cannot be assessed (e.g., previously removed).Stage N0 indicates no regional lymph node metastasis. Stage N1 indicatesmetastasis to movable ipsilateral axillary lymph nodes. Stage N2indicates metastasis to ipsilateral axillary lymph nodes fixed to oneanother or to other structures. Stage N3 indicates metastasis toipsilateral internal mammary lymph nodes. Id.

[0011] Stage determination has potential prognostic value and providescriteria for designing optimal therapy. Simpson et al., 18 J. Clin.Oncology 2059 (2000). Generally, pathological staging of breast canceris preferable to clinical staging because the former gives a moreaccurate prognosis. However, clinical staging would be preferred if itwere as accurate as pathological staging because it does not depend onan invasive procedure to obtain tissue for pathological evaluation.Staging of breast cancer would be improved by detecting new markers incells, tissues, or bodily fluids which could differentiate betweendifferent stages of invasion. Progress in this field will allow morerapid and reliable method for treating breast cancer patients.

[0012] Treatment of breast cancer is generally decided after an accuratestaging of the primary tumor. Primary treatment options include breastconserving therapy (lumpectomy, breast irradiation, and surgical stagingof the axilla), and modified radical mastectomy. Additional treatmentsinclude chemotherapy, regional irradiation, and, in extreme cases,terminating estrogen production by ovarian ablation.

[0013] Until recently, the customary treatment for all breast cancer wasmastectomy. Fonseca et al., 127 Annals of Internal Medicine 1013 (1997).However, recent data indicate that less radical procedures may beequally effective, in terms of survival, for early stage breast cancer.Fisher et al., 16 J. of Clinical Oncology 441 (1998). The treatmentoptions for a patient with early stage breast cancer (i.e., stage Tis)may be breast-sparing surgery followed by localized radiation therapy atthe breast. Alternatively, mastectomy optionally coupled with radiationor breast reconstruction may be employed. These treatment methods areequally effective in the early stages of breast cancer.

[0014] Patients with stage I and stage II breast cancer require surgerywith chemotherapy and/or hormonal therapy. Surgery is of limited use inStage III and stage IV patients. Thus, these patients are bettercandidates for chemotherapy and radiation therapy with surgery limitedto biopsy to permit initial staging or subsequent restaging becausecancer is rarely curative at this stage of the disease. AJCC CancerStaging Handbook 84, ¶. 164-65 (Irvin D. Fleming et al. eds., 5^(th) ed.1998).

[0015] In an effort to provide more treatment options to patients,efforts are underway to define an earlier stage of breast cancer withlow recurrence which could be treated with lumpectomy withoutpostoperative radiation treatment. While a number of attempts have beenmade to classify early stage breast cancer, no consensus recommendationon postoperative radiation treatment has been obtained from thesestudies. Page et al., 75 Cancer 1219 (1995); Fisher et al., 75 Cancer1223 (1995); Silverstein et al., 77 Cancer 2267(1996).

[0016] As discussed above, each of the methods for diagnosing andstaging breast cancer is limited by the technology employed.Accordingly, there is need for sensitive molecular and cellular markersfor the detection of breast cancer. There is a need for molecularmarkers for the accurate staging, including clinical and pathologicalstaging, of breast cancers to optimize treatment methods. Finally, thereis a need for sensitive molecular and cellular markers to monitor theprogress of cancer treatments, including markers that can detectrecurrence of breast cancers following remission.

[0017] Other objects, features, advantages and aspects of the presentinvention will become apparent to those of skill in the art from thefollowing description. It should be understood, however, that thefollowing description and the specific examples, while indicatingpreferred embodiments of the invention, are given by way of illustrationonly. Various changes and modifications within the spirit and scope ofthe disclosed invention will become readily apparent to those skilled inthe art from reading the following description and from reading theother parts of the present disclosure.

SUMMARY OF THE INVENTION

[0018] The present invention solves these and other needs in the art byproviding nucleic acid molecules and polypeptides as well as antibodies,agonists and antagonists, thereto that may be used to identify,diagnose, monitor, stage, image and treat breast cancer andnon-cancerous disease states in breast; identify and monitor breasttissue; and identify and design agonists and antagonists of polypeptidesof the invention. The invention also provides gene therapy, methods forproducing transgenic animals and cells, and methods for producingengineered breast tissue for treatment and research.

[0019] Accordingly, one object of the invention is to provide nucleicacid molecules that are specific to breast cells and/or breast tissue.These breast specific nucleic acids (BSNAs) may be a naturally-occurringcDNA, genomic DNA, RNA, or a fragment of one of these nucleic acids, ormay be a non-naturally-occurring nucleic acid molecule. If the BSNA isgenomic DNA, then the BSNA is a breast specific gene (BSG). In apreferred embodiment, the nucleic acid molecule encodes a polypeptidethat is specific to breast. In a more preferred embodiment, the nucleicacid molecule encodes a polypeptide that comprises an amino acidsequence of SEQ ID NO: 160 through 282. In another highly preferredembodiment, the nucleic acid molecule comprises a nucleic acid sequenceof SEQ ID NO: 1 through 159. By nucleic acid molecule, it is also meantto be inclusive of sequences that selectively hybridize or exhibitsubstantial sequence similarity to a nucleic acid molecule encoding aBSP, or that selectively hybridize or exhibit substantial sequencesimilarity to a BSNA, as well as allelic variants of a nucleic acidmolecule encoding a BSP, and allelic variants of a BSNA. Nucleic acidmolecules comprising a part of a nucleic acid sequence that encodes aBSP or that comprises a part of a nucleic acid sequence of a BSNA arealso provided.

[0020] A related object of the present invention is to provide a nucleicacid molecule comprising one or more expression control sequencescontrolling the transcription and/or translation of all or a part of aBSNA. In a preferred embodiment, the nucleic acid molecule comprises oneor more expression control sequences controlling the transcriptionand/or translation of a nucleic acid molecule that encodes all or afragment of a BSP.

[0021] Another object of the invention is to provide vectors and/or hostcells comprising a nucleic acid molecule of the instant invention. In apreferred embodiment, the nucleic acid molecule encodes all or afragment of a BSP. In another preferred embodiment, the nucleic acidmolecule comprises all or a part of a BSNA.

[0022] Another object of the invention is to provided methods for usingthe vectors and host cells comprising a nucleic acid molecule of theinstant invention to recombinantly produce polypeptides of theinvention.

[0023] Another object of the invention is to provide a polypeptideencoded by a nucleic acid molecule of the invention. In a preferredembodiment, the polypeptide is a BSP. The polypeptide may compriseeither a fragment or a full-length protein as well as a mutant protein(mutein), fusion protein, homologous protein or a polypeptide encoded byan allelic variant of a BSP.

[0024] Another object of the invention is to provide an antibody thatspecifically binds to a polypeptide of the instant invention.

[0025] Another object of the invention is to provide agonists andantagonists of the nucleic acid molecules and polypeptides of theinstant invention.

[0026] Another object of the invention is to provide methods for usingthe nucleic acid molecules to detect or amplify nucleic acid moleculesthat have similar or identical nucleic acid sequences compared to thenucleic acid molecules described herein. In a preferred embodiment, theinvention provides methods of using the nucleic acid molecules of theinvention for identifying, diagnosing, monitoring, staging, imaging andtreating breast cancer and non-cancerous disease states in breast. Inanother preferred embodiment, the invention provides methods of usingthe nucleic acid molecules of the invention for identifying and/ormonitoring breast tissue. The nucleic acid molecules of the instantinvention may also be used in gene therapy, for producing transgenicanimals and cells, and for producing engineered breast tissue fortreatment and research.

[0027] The polypeptides and/or antibodies of the instant invention mayalso be used to identify, diagnose, monitor, stage, image and treatbreast cancer and non-cancerous disease states in breast. The inventionprovides methods of using the polypeptides of the invention to identifyand/or monitor breast tissue, and to produce engineered breast tissue.

[0028] The agonists and antagonists of the instant invention may be usedto treat breast cancer and non-cancerous disease states in breast and toproduce engineered breast tissue.

[0029] Yet another object of the invention is to provide a computerreadable means of storing the nucleic acid and amino acid sequences ofthe invention. The records of the computer readable means can beaccessed for reading and displaying of sequences for comparison,alignment and ordering of the sequences of the invention to othersequences.

DETAILED DESCRIPTION OF THE INVENTION

[0030] Definitions and General Techniques

[0031] Unless otherwise defined herein, scientific and technical termsused in connection with the present invention shall have the meaningsthat are commonly understood by those of ordinary skill in the art.Further, unless otherwise required by context, singular terms shallinclude pluralities and plural terms shall include the singular.Generally, nomenclatures used in connection with, and techniques of,cell and tissue culture, molecular biology, immunology, microbiology,genetics and protein and nucleic acid chemistry and hybridizationdescribed herein are those well-known and commonly used in the art. Themethods and techniques of the present invention are generally performedaccording to conventional methods well-known in the art and as describedin various general and more specific references that are cited anddiscussed throughout the present specification unless otherwiseindicated. See, e.g., Sambrook et al., Molecular Cloning: A LaboratoryManual, 2d ed., Cold Spring Harbor Laboratory Press (1989) and Sambrooket al., Molecular Cloning: A Laboratory Manual, 3d ed., Cold SpringHarbor Press (2001); Ausubel et al., Current Protocols in MolecularBiology, Greene Publishing Associates (1992, and Supplements to 2000);Ausubel et al., Short Protocols in Molecular Biology: A Compendium ofMethods from Current Protocols in Molecular Biology—4^(th) Ed., Wiley &Sons (1999); Harlow and Lane, Antibodies: A Laboratory Manual, ColdSpring Harbor Laboratory Press (1990); and Harlow and Lane, UsingAntibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press(1999); each of which is incorporated herein by reference in itsentirety.

[0032] Enzymatic reactions and purification techniques are performedaccording to manufacturer's specifications, as commonly accomplished inthe art or as described herein. The nomenclatures used in connectionwith, and the laboratory procedures and techniques of, analyticalchemistry, synthetic organic chemistry, and medicinal and pharmaceuticalchemistry described herein are those well-known and commonly used in theart. Standard techniques are used for chemical syntheses, chemicalanalyses, pharmaceutical preparation, formulation, and delivery, andtreatment of patients.

[0033] The following terms, unless otherwise indicated, shall beunderstood to have the following meanings:

[0034] A “nucleic acid molecule” of this invention refers to a polymericform of nucleotides and includes both sense and antisense strands ofRNA, cDNA, genomic DNA, and synthetic forms and mixed polymers of theabove. A nucleotide refers to a ribonucleotide, deoxynucleotide or amodified form of either type of nucleotide. A “nucleic acid molecule” asused herein is synonymous with “nucleic acid” and “polynucleotide.” Theterm “nucleic acid molecule” usually refers to a molecule of at least 10bases in length, unless otherwise specified. The term includes single-and double-stranded forms of DNA. In addition, a polynucleotide mayinclude either or both naturally-occurring and modified nucleotideslinked together by naturally-occurring and/or non-naturally occurringnucleotide linkages.

[0035] The nucleic acid molecules may be modified chemically orbiochemically or may contain non-natural or derivatized nucleotidebases, as will be readily appreciated by those of skill in the art. Suchmodifications include, for example, labels, methylation, substitution ofone or more of the naturally occurring nucleotides with an analog,internucleotide modifications such as uncharged linkages (e.g., methylphosphonates, phosphotriesters, phosphoramidates, carbamates, etc.),charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.),pendent moieties (e.g., polypeptides), intercalators (e.g., acridine,psoralen, etc.), chelators, alkylators, and modified linkages (e.g.,alpha anomeric nucleic acids, etc.) The term “nucleic acid molecule”also includes any topological conformation, including single-stranded,double-stranded, partially duplexed, triplexed, hairpinned, circular andpadlocked conformations. Also included are synthetic molecules thatmimic polynucleotides in their ability to bind to a designated sequencevia hydrogcn bonding and other chemical interactions. Such molecules areknown in the art and include, for example, those in which peptidelinkages substitute for phosphate linkages in the backbone of themolecule.

[0036] A “gene” is defined as a nucleic acid molecule that comprises anucleic acid sequence that encodes a polypeptide and the expressioncontrol sequences that surround the nucleic acid sequence that encodesthe polypeptide. For instance, a gene may comprise a promoter, one ormore enhancers, a nucleic acid sequence that encodes a polypeptide,downstream regulatory sequences and, possibly, other nucleic acidsequences involved in regulation of the expression of an RNA. As iswell-known in the art, eukaryotic genes usually contain both exons andintrons. The term “exon” refers to a nucleic acid sequence found ingenomic DNA that is bioinformatically predicted and/or experimentallyconfirmed to contribute a contiguous sequence to a mature mRNAtranscript. The term “intron” refers to a nucleic acid sequence found ingenomic DNA that is predicted and/or confirmed to not contribute to amature mRNA transcript, but rather to be “spliced out” during processingof the transcript.

[0037] A nucleic acid molecule or polypeptide is “derived” from aparticular species if the nucleic acid molecule or polypeptide has beenisolated from the particular species, or if the nucleic acid molecule orpolypeptide is homologous to a nucleic acid molecule or polypeptideisolated from a particular species.

[0038] An “isolated” or “substantially pure” nucleic acid orpolynucleotide (e.g., an RNA, DNA or a mixed polymer) is one which issubstantially separated from other cellular components that naturallyaccompany the native polynucleotide in its natural host cell, e.g.,ribosomes, polymerases, or genomic sequences with which it is naturallyassociated. The term embraces a nucleic acid or polynucleotide that (1)has been removed from its naturally occurring environment, (2) is notassociated with all or a portion of a polynucleotide in which the“isolated polynucleotide” is found in nature, (3) is operatively linkedto a polynucleotide which it is not linked to in nature, (4) does notoccur in nature as part of a larger sequence or (5) includes nucleotidesor internucleoside bonds that are not found in nature. The term“isolated” or “substantially pure” also can be used in reference torecombinant or cloned DNA isolates, chemically synthesizedpolynucleotide analogs, or polynucleotide analogs that are biologicallysynthesized by heterologous systems. The term “isolated nucleic acidmolecule” includes nucleic acid molecules that are integrated into ahost cell chromosome at a heterologous site, recombinant fusions of anative fragment to a heterologous sequence, recombinant vectors presentas episomes or as integrated into a host cell chromosome.

[0039] A “part” of a nucleic acid molecule refers to a nucleic acidmolecule that comprises a partial contiguous sequence of at least 10bases of the reference nucleic acid molecule. Preferably, a partcomprises at least 15 to 20 bases of a reference nucleic acid molecule.In theory, a nucleic acid sequence of 17 nucleotides is of sufficientlength to occur at random less frequently than once in the threegigabase human genome, and thus to provide a nucleic acid probe that canuniquely identify the reference sequence in a nucleic acid mixture ofgenomic complexity. A preferred part is one that comprises a nucleicacid sequence that can encode at least 6 contiguous amino acid sequences(fragments of at least 18 nucleotides) because they are useful indirecting the expression or synthesis of peptides that are useful inmapping the epitopes of the polypeptide encoded by the reference nucleicacid. See, e.g., Geysen et al., Proc. Natl. Acad. Sci. USA 81:3998-4002(1984); and U.S. Pat. Nos. 4,708,871 and 5,595,915, the disclosures ofwhich are incorporated herein by reference in their entireties. A partmay also comprise at least 25, 30, 35 or 40 nucleotides of a referencenucleic acid molecule, or at least 50, 60, 70, 80, 90, 100, 150, 200,250, 300, 350, 400 or 500 nucleotides of a reference nucleic acidmolecule. A part of a nucleic acid molecule may comprise no othernucleic acid sequences. Alternatively, a part of a nucleic acid maycomprise other nucleic acid sequences from other nucleic acid molecules.

[0040] The term “oligonucleotide” refers to a nucleic acid moleculegenerally comprising a length of 200 bases or fewer. The term oftenrefers to single-stranded deoxyribonucleotides, but it can refer as wellto single- or double-stranded ribonucleotides, RNA:DNA hybrids anddouble-stranded DNAs, among others. Preferably, oligonucleotides are 10to 60 bases in length and most preferably 12, 13, 14, 15, 16, 17, 18, 19or 20 bases in length. Other preferred oligonucleotides are 25, 30, 35,40, 45, 50, 55 or 60 bases in length. Oligonucleotides may besingle-stranded, e.g. for use as probes or primers, or may bedouble-stranded, e.g. for use in the construction of a mutant gene.Oligonucleotides of the invention can be either sense or antisenseoligonucleotides. An oligonucleotide can be derivatized or modified asdiscussed above for nucleic acid molecules.

[0041] Oligonuclcotidcs, such as single-stranded DNA probeoligonucleotides, often are synthesized by chemical methods, such asthose implemented on automated oligonucleotide synthesizers. However,oligonucleotides can be made by a variety of other methods, including invitro recombinant DNA-mediated techniques and by expression of DNAs incells and organisms. Initially, chemically synthesized DNAs typicallyare obtained without a 5′ phosphate. The 5′ ends of sucholigonucleotides are not substrates for phosphodiester bond formation byligation reactions that employ DNA ligases typically used to formrecombinant DNA molecules. Where ligation of such oligonucleotides isdesired, a phosphate can be added by standard techniques, such as thosethat employ a kinase and ATP. The 3′ end of a chemically synthesizedoligonucleotide generally has a free hydroxyl group and, in the presenceof a ligase, such as T4 DNA ligase, readily will form a phosphodiesterbond with a 5′ phosphate of another polynucleotide, such as anotheroligonucleotide. As is well-known, this reaction can be preventedselectively, where desired, by removing the 5′ phosphates of the otherpolynucleotide(s) prior to ligation.

[0042] The term “naturally-occurring nucleotide” referred to hereinincludes naturally-occurring deoxyribonucleotides and ribonucleotides.The term “modified nucleotides” referred to herein includes nucleotideswith modified or substituted sugar groups and the like. The term“nucleotide linkages” referred to herein includes nucleotides linkagessuch as phosphorothioate, phosphorodithioate, phosphoroselenoate,phosphorodiselenoate, phosphoroanilothioate, phoshoraniladate,phosphoroamidate, and the like. See e.g., LaPlanche et al. Nucl. AcidsRes. 14:9081-9093 (1986); Stein et al. Nucl. Acids Res. 16:3209-3221(1988); Zon et al. Anti-Cancer Drug Design 6:539-568 (1991); Zon et al.,in Eckstein (ed.) Oligonucleotides and Analogues: A Practical Approach,pp. 87-108, Oxford University Press (1991); U.S. Pat. No. 5,151,510;Uhlmann and Peyman Chemical Reviews 90:543 (1990), the disclosures ofwhich are hereby incorporated by reference.

[0043] Unless specified otherwise, the left hand end of a polynucleotidesequence in sense orientation is the 5′ end and the right hand end ofthe sequence is the 3′ end. In addition, the left hand direction of apolynucleotide sequence in sense orientation is referred to as the 5′direction, while the right hand direction of the polynucleotide sequenceis referred to as the 3′ direction. Further, unless otherwise indicated,each nuclcotide sequence is set forth herein as a sequence ofdeoxyribonucleotides. It is intended, however, that the given sequencebe interpreted as would be appropriate to the polynucleotidecomposition: for example, if the isolated nucleic acid is composed ofRNA, the given sequence intends ribonucleotides, with uridinesubstituted for thymidine.

[0044] The term “allelic variant” refers to one of two or morealternative naturally-occurring forms of a gene, wherein each genepossesses a unique nucleotide sequence. In a preferred embodiment,different alleles of a given gene have similar or identical biologicalproperties.

[0045] The term “percent sequence identity” in the context of nucleicacid sequences refers to the residues in two sequences which are thesame when aligned for maximum correspondence. The length of sequenceidentity comparison may be over a stretch of at least about ninenucleotides, usually at least about 20 nucleotides, more usually atleast about 24 nuclcotides, typically at least about 28 nuclcotides,more typically at least about 32 nucleotides, and preferably at leastabout 36 or more nucleotides. There are a number of different algorithmsknown in the art which can be used to measure nucleotide sequenceidentity. For instance, polynucleotide sequences can be compared usingFASTA, Gap or Bestfit, which are programs in Wisconsin Package Version10.0, Genetics Computer Group (GCG), Madison, Wis. FASTA, whichincludes, e.g., the programs FASTA2 and FASTA3, provides alignments andpercent sequence identity of the regions of the best overlap between thequery and search sequences (Pearson, Methods Enzymol. 183: 63-98 (1990);Pearson, Methods Mol. Biol. 132: 185-219 (2000); Pearson, MethodsEnzymol. 266: 227-258 (1996); Pearson, J. Mol. Biol. 276: 71-84 (1998);herein incorporated by reference). Unless otherwise specified, defaultparameters for a particular program or algorithm are used. For instance,percent sequence identity between nucleic acid sequences can bedetermined using FASTA with its default parameters (a word size of 6 andthe NOPAM factor for the scoring matrix) or using Gap with its defaultparameters as provided in GCG Version 6.1, herein incorporated byreference.

[0046] A reference to a nucleic acid sequence encompasses its complementunless otherwise specified. Thus, a reference to a nucleic acid moleculehaving a particular sequence should be understood to encompass itscomplementary strand, with its complementary sequence. The complementarystrand is also useful, e.g., for antisense therapy, hybridization probesand PCR primers.

[0047] In the molecular biology art, researchers use the terms “percentsequence identity”, “percent sequence similarity” and “percent sequencehomology” interchangeably. In this application, these terms shall havethe same meaning with respect to nucleic acid sequences only.

[0048] The term “substantial similarity” or “substantial sequencesimilarity,” when referring to a nucleic acid or fragment thereof,indicates that, when optimally aligned with appropriate nucleotideinsertions or deletions with another nucleic acid (or its complementarystrand), there is nucleotide sequence identity in at least about 50%,more preferably 60% of the nucleotide bases, usually at least about 70%,more usually at least about 80%, preferably at least about 90%, and morepreferably at least about 95-98% of the nucleotide bases, as measured byany well-known algorithm of sequence identity, such as FASTA, BLAST orGap, as discussed above.

[0049] Alternatively, substantial similarity exists when a nucleic acidor fragment thereof hybridizes to another nucleic acid, to a strand ofanother nucleic acid, or to the complementary strand thereof, underselective hybridization conditions. Typically, selective hybridizationwill occur when there is at least about 55% sequence identity,preferably at least about 65%, more preferably at least about 75%, andmost preferably at least about 90% sequence identity, over a stretch ofat least about 14 nucleotides, more preferably at least 17 nucleotides,even more preferably at least 20, 25, 30, 35, 40, 50, 60, 70, 80, 90 or100 nucleotides.

[0050] Nucleic acid hybridization will be affected by such conditions assalt concentration, temperature, solvents, the base composition of thehybridizing species, length of the complementary regions, and the numberof nucleotide base mismatches between the hybridizing nucleic acids, aswill be readily appreciated by those skilled in the art. “Stringenthybridization conditions” and “stringent wash conditions” in the contextof nucleic acid hybridization experiments depend upon a number ofdifferent physical parameters. The most important parameters includetemperature of hybridization, base composition of the nucleic acids,salt concentration and length of the nucleic acid. One having ordinaryskill in the art knows how to vary these parameters to achieve aparticular stringency of hybridization. In general, “stringenthybridization” is performed at about 25° C. below the thermal meltingpoint (T_(m)) for the specific DNA hybrid under a particular set ofconditions. “Stringent washing” is performed at temperatures about 5° C.lower than the T_(m) for the specific DNA hybrid under a particular setof conditions. The T_(m) is the temperature at which 50% of the targetsequence hybridizes to a perfectly matched probe. See Sambrook (1989),supra, p.9.51, hereby incorporated by reference.

[0051] The T_(m) for a particular DNA-DNA hybrid can be estimated by theformula:

T_(m)=81.5° C.+16.6(log₁₀[Na⁺])+0.41 (fraction G+C)−0.63 (%formamide)−(600/1)

[0052] where 1 is the length of the hybrid in base pairs.

[0053] The T_(m) for a particular RNA-RNA hybrid can be estimated by theformula:

T_(m)=79.8° C.+18.5(log₁₀[Na⁺])+0.58 (fraction G+C)+11.8 (fractionG+C)²−0.35 (% formamide)−(820/1).

[0054] The T_(m) for a particular RNA-DNA hybrid can be estimated by theformula:

T_(m)=79.8° C.+18.5(log₁₀[Na⁺])+0.58 (fraction G+C)+11.8 (fractionG+C)²−0.50 (% formamide)−(820/1).

[0055] In general, the T_(m) decreases by 1-1.5° C. for each 1% ofmismatch between two nucleic acid sequences. Thus, one having ordinaryskill in the art can alter hybridization and/or washing conditions toobtain sequences that have higher or lower degrees of sequence identityto the target nucleic acid. For instance, to obtain hybridizing nucleicacids that contain up to 10% mismatch from the target nucleic acidsequence, 10-15° C. would be subtracted from the calculated T_(m) of aperfectly matched hybrid, and then the hybridization and washingtemperatures adjusted accordingly. Probe sequences may also hybridizespecifically to duplex DNA under certain conditions to form triplex orother higher order DNA complexes. The preparation of such probes andsuitable hybridization conditions are well-known in the art.

[0056] An example of stringent hybridization conditions forhybridization of complementary nucleic acid sequences having more than100 complementary residues on a filter in a Southern or Northern blot orfor screening a library is 50% formamide/6×SSC at 42° C. for at leastten hours and preferably overnight (approximately 16 hours). Anotherexample of stringent hybridization conditions is 6×SSC at 68° C. withoutformamide for at least ten hours and preferably overnight. An example ofmoderate stringency hybridization conditions is 6×SSC at 55° C. withoutformamide for at least ten hours and preferably overnight. An example oflow stringency hybridization conditions for hybridization ofcomplementary nucleic acid sequences having more than 100 complementaryresidues on a filter in a Southern or Northern blot or for screening alibrary is 6×SSC at 42° C. for at least ten hours. Hybridizationconditions to identify nucleic acid sequences that are similar but notidentical can be identified by experimentally changing the hybridizationtemperature from 68° C. to 42° C. while keeping the salt concentrationconstant (6×SSC), or keeping the hybridization temperature and saltconcentration constant (e.g. 42° C. and 6×SSC) and varying the formamideconcentration from 50% to 0%. Hybridization buffers may also includeblocking agents to lower background. These agents are well-known in theart. See Sambrook et al. (1989), supra, pages 8.46 and 9.46-9.58, hereinincorporated by reference. See also Ausubel (1992), supra, Ausubel(1999), supra, and Sambrook (2001), supra.

[0057] Wash conditions also can be altered to change stringencyconditions. An example of stringent wash conditions is a 0.2×SSC wash at65° C. for 15 minutes (see Sambrook (1989), supra, for SSC buffer).Often the high stringency wash is preceded by a low stringency wash toremove excess probe. An exemplary medium stringency wash for duplex DNAof more than 100 base pairs is 1×SSC at 45° C. for 15 minutes. Anexemplary low stringency wash for such a duplex is 4×SSC at 40° C. for15 minutes. In general, signal-to-noise ratio of 2× or higher than thatobserved for an unrelated probe in the particular hybridization assayindicates detection of a specific hybridization.

[0058] As defined herein, nucleic acid molecules that do not hybridizeto each other under stringent conditions are still substantially similarto one another if they encode polypeptides that are substantiallyidentical to each other. This occurs, for example, when a nucleic acidmolecule is created synthetically or recombinantly using high codondegeneracy as permitted by the redundancy of the genetic code.

[0059] Hybridization conditions for nucleic acid molecules that areshorter than 100 nucleotides in length (e.g., for oligonucleotideprobes) may be calculated by the formula:

T_(m)=81.5° C.+16.6(log₁₀[Na⁺])+0.41(fraction G+C)−(600/N),

[0060] wherein N is change length and the [Na⁺] is 1 M or less. SeeSambrook (1989), supra, p. 11.46. For hybridization of probes shorterthan 100 nucleotides, hybridization is usually performed under stringentconditions (5-10° C. below the T_(m)) using high concentrations (0.1-1.0pmol/ml) of probe. Id. at p. 11.45. Determination of hybridization usingmismatched probes, pools of degenerate probes or “guessmers,” as well ashybridization solutions and methods for empirically determininghybridization conditions are well-known in the art. See, e.g., Ausubel(1999), supra; Sambrook (1989), supra, pp. 11.45-11.57.

[0061] The term “digestion” or “digestion of DNA” refers to catalyticcleavage of the DNA with a restriction enzyme that acts only at certainsequences in the DNA. The various restriction enzymes referred to hereinare commercially available and their reaction conditions, cofactors andother requirements for use are known and routine to the skilled artisan.For analytical purposes, typically, 1 μg of plasmid or DNA fragment isdigested with about 2 units of enzyme in about 20 μl of reaction buffer.For the purpose of isolating DNA fragments for plasmid construction,typically 5 to 50 μg of DNA are digested with 20 to 250 units of enzymein proportionately larger volumes. Appropriate buffers and substrateamounts for particular restriction enzymes are described in standardlaboratory manuals, such as those referenced below, and they arespecified by commercial suppliers. Incubation times of about 1 hour at37° C. are ordinarily used, but conditions may vary in accordance withstandard procedures, the supplier's instructions and the particulars ofthe reaction. After digestion, reactions may be analyzed, and fragmentsmay be purified by electrophoresis through an agarose or polyacrylamidegel, using well-known methods that are routine for those skilled in theart.

[0062] The term “ligation” refers to the process of formingphosphodiester bonds between two or more polynucleotides, which mostoften are double-stranded DNAS. Techniques for ligation are well-knownto the art and protocols for ligation are described in standardlaboratory manuals and references, such as, e.g., Sambrook (1989),supra.

[0063] Genome-derived “single exon probes,” are probes that comprise atleast part of an exon (“reference exon”) and can hybridize detectablyunder high stringency conditions to transcript-derived nucleic acidsthat include the reference exon but do not hybridize detectably underhigh stringency conditions to nucleic acids that lack the referenceexon. Single exon probes typically further comprise, contiguous to afirst end of the exon portion, a first intronic and/or intergenicsequence that is identically contiguous to the exon in the genome, andmay contain a second intronic and/or intergenic sequence that isidentically contiguous to the exon in the genome. The minimum length ofgenome-derived single exon probes is defined by the requirement that theexonic portion be of sufficient length to hybridize under highstringency conditions to transcript-derived nucleic acids, as discussedabove. The maximum length of genome-derived single exon probes isdefined by the requirement that the probes contain portions of no morethan one exon. The single exon probes may contain priming sequences notfound in contiguity with the rest of the probe sequence in the genome,which priming sequences are useful for PCR and other amplification-basedtechnologies.

[0064] The term “microarray” or “nucleic acid microarray” refers to asubstrate-bound collection of plural nucleic acids, hybridization toeach of the plurality of bound nucleic acids being separatelydetectable. The substrate can be solid or porous, planar or non-planar,unitary or distributed. Microarrays or nucleic acid microarrays includeall the devices so called in Schena (ed.), DNA Microarrays: A PracticalApproach (Practical Approach Series), Oxford University Press (1999);Nature Genet. 21 (1)(suppl.): 1-60 (1999); Schena (ed.), MicroarrayBiochip: Tools and Technology, Eaton Publishing Company/BioTechniquesBooks Division (2000). These microarrays include substrate-boundcollections of plural nucleic acids in which the plurality of nucleicacids are disposed on a plurality of beads, rather than on a unitaryplanar substrate, as is described, inter alia, in Brenner et al., Proc.Natl. Acad. Sci. USA 97(4):1665-1670 (2000).

[0065] The term “mutated” when applied to nucleic acid molecules meansthat nucleotides in the nucleic acid sequence of the nucleic acidmolecule may be inserted, deleted or changed compared to a referencenucleic acid sequence. A single alteration may be made at a locus (apoint mutation) or multiple nucleotides may be inserted, deleted orchanged at a single locus. In addition, one or more alterations may bemade at any number of loci within a nucleic acid sequence. In apreferred embodiment, the nucleic acid molecule comprises the wild typenucleic acid sequence encoding a BSP or is a BSNA. The nucleic acidmolecule may be mutated by any method known in the art including thosemutagenesis techniques described infra.

[0066] The term “error-prone PCR” refers to a process for performing PCRunder conditions where the copying fidelity of the DNA polymerase islow, such that a high rate of point mutations is obtained along theentire length of the PCR product. See, e.g., Leung et al., Technique 1:11-15 (1989) and Caldwell et al., PCR Methods Applic. 2: 28-33 (1992).

[0067] The term “oligonucleotide-directed mutagenesis” refers to aprocess which enables the generation of site-specific mutations in anycloned DNA segment of interest. See, e.g., Reidhaar-Olson et al.,Science 241: 53-57 (1988).

[0068] The term “assembly PCR” refers to a process which involves theassembly of a PCR product from a mixture of small DNA fragments. A largenumber of different PCR reactions occur in parallel in the same vial,with the products of one reaction priming the products of anotherreaction.

[0069] The term “sexual PCR mutagenesis” or “DNA shuffling” refers to amethod of error-prone PCR coupled with forced homologous recombinationbetween DNA molecules of different but highly related DNA sequence invitro, caused by random fragmentation of the DNA molecule based onsequence similarity, followed by fixation of the crossover by primerextension in an error-prone PCR reaction. See, e.g., Stemmer, Proc.Natl. Acad. Sci. U.S.A. 91: 10747-10751 (1994). DNA shuffling can becarried out between several related genes (“Family shuffling”).

[0070] The term “in vivo mutagenesis” refers to a process of generatingrandom mutations in any cloned DNA of interest which involves thepropagation of the DNA in a strain of bacteria such as E. coli thatcarries mutations in onc or morc of the DNA repair pathways. These“mutator” strains have a higher random mutation rate than that of awild-type parent. Propagating the DNA in a mutator strain willeventually generate random mutations within the DNA.

[0071] The term “cassette mutagenesis” refers to any process forreplacing a small region of a double-stranded DNA molecule with asynthetic oligonucleotide “cassette” that differs from the nativesequence. The oligonucleotide often contains completely and/or partiallyrandomized native sequence.

[0072] The term “recursive ensemble mutagenesis” refers to an algorithmfor protein engineering (protein mutagenesis) developed to producediverse populations of phenotypically related mutants whose membersdiffer in amino acid sequence. This method uses a feedback mechanism tocontrol successive rounds of combinatorial cassette mutagenesis. See,e.g., Arkin et al., Proc. Natl. Acad. Sci. U.S.A. 89: 7811-7815 (1992).

[0073] The term “exponential ensemble mutagenesis” refers to a processfor generating combinatorial libraries with a high pcrcentagc of uniqueand functional mutants, wherein small groups of residues are randomizedin parallel to identify, at each altered position, amino acids whichlead to functional proteins. See, e.g., Delegrave et al., BiotechnologyResearch 11: 1548-1552 (1993); Arnold, Current Opinion in Biotechnology4: 450-455 (1993). Each of the references mentioned above are herebyincorporated by reference in its entirety.

[0074] “Operatively linked” expression control sequences refers to alinkage in which the expression control sequence is contiguous with thegene of interest to control the gene of interest, as well as expressioncontrol sequences that act in trans or at a distance to control the geneof interest.

[0075] The term “expression control sequence” as used herein refers topolynucleotide sequences which are necessary to affect the expression ofcoding sequences to which they are operatively linked. Expressioncontrol sequences are sequences which control the transcription,post-transcriptional events and translation of nucleic acid sequences.Expression control sequences include appropriate transcriptioninitiation, termination, promoter and enhancer sequences; efficient RNAprocessing signals such as splicing and polyadenylation signals;sequences that stabilize cytoplasmic mRNA; sequences that enhancetranslation efficiency (e.g., ribosome binding sites); sequences thatenhance protein stability; and when desired, sequences that enhanceprotein secretion. The nature of such control sequences differsdepending upon the host organism; in prokaryotes, such control sequencesgenerally include the promoter, ribosomal binding site, andtranscription termination sequence. The term “control sequences” isintended to include, at a minimum, all components whose presence isessential for expression, and can also include additional componentswhose presence is advantageous, for example, leader sequences and fusionpartner sequences.

[0076] The term “vector,” as used herein, is intended to refer to anucleic acid molecule capable of transporting another nucleic acid towhich it has been linked. One type of vector is a “plasmid”, whichrefers to a circular double-stranded DNA loop into which additional DNAsegments may be ligated. Other vectors include cosmids, bacterialartificial chromosomes (BAC) and yeast artificial chromosomes (YAC).Another type of vector is a viral vector, wherein additional DNAsegments may be ligated into the viral genome. Viral vectors that infectbacterial cells are referred to as bacteriophages. Certain vectors arecapable of autonomous replication in a host cell into which they areintroduced (e.g., bacterial vectors having a bacterial origin ofreplication). Other vectors can be integrated into the genome of a hostcell upon introduction into the host cell, and thereby are replicatedalong with the host genome. Moreover, certain vectors are capable ofdirecting the expression of genes to which they are operatively linked.Such vectors are referred to herein as “recombinant expression vectors”(or simply, “expression vectors”). In general, expression vectors ofutility in recombinant DNA techniques are often in the form of plasmids.In the present specification, “plasmid” and “vector” may be usedinterchangeably as the plasmid is the most commonly used form of vector.However, the invention is intended to include other forms of expressionvectors that serve equivalent functions.

[0077] The term “recombinant host cell” (or simply “host cell”), as usedherein, is intended to refer to a cell into which an expression vectorhas been introduced. It should be understood that such terms areintended to refer not only to the particular subject cell but to theprogeny of such a cell. Because certain modifications may occur insucceeding generations due to either mutation or environmentalinfluences, such progeny may not, in fact, be identical to the parentcell, but are still included within the scope of the term “host cell” asused herein.

[0078] As used herein, the phrase “open reading frame” and theequivalent acronym “ORF” refer to that portion of a transcript-derivednucleic acid that can be translated in its entirety into a sequence ofcontiguous amino acids. As so defined, an ORF has length, measured innucleotides, exactly divisible by 3. As so defined, an ORF need notencode the entirety of a natural protein.

[0079] As used herein, the phrase “ORF-encoded peptide” refers to thepredicted or actual translation of an ORF.

[0080] As used herein, the phrase “degenerate variant” of a referencenucleic acid sequence intends all nucleic acid sequences that can bedirectly translated, using the standard genetic code, to provide anamino acid sequence identical to that translated from the referencenucleic acid sequence.

[0081] The term “polypeptide” encompasses both naturally-occurring andnon-naturally-occurring proteins and polypeptides, polypeptide fragmentsand polypeptide mutants, derivatives and analogs. A polypeptide may bemonomeric or polymeric. Further, a polypeptide may comprise a number ofdiffract modules within a single polypeptide each of which has one ormore distinct activities. A preferred polypeptide in accordance with theinvention comprises a BSP encoded by a nucleic acid molecule of theinstant invention, as well as a fragment, mutant, analog and derivativethereof.

[0082] The term “isolated protein” or “isolated polypeptide” is aprotein or polypeptide that by virtue of its origin or source ofderivation (1) is not associated with naturally associated componentsthat accompany it in its native state, (2) is free of other proteinsfrom the same species (3) is expressed by a cell from a differentspecies, or (4) does not occur in nature. Thus, a polypeptide that ischemically synthesized or synthesized in a cellular system differentfrom the cell from which it naturally originates will be “isolated” fromits naturally associated components. A polypeptide or protein may alsobe rendered substantially free of naturally associated components byisolation, using protein purification techniques well-known in the art.

[0083] A protein or polypeptide is “substantially pure,” “substantiallyhomogeneous” or “substantially purified” when at least about 60% to 75%of a sample exhibits a single species of polypeptide. The polypeptide orprotein may be monomeric or multimeric. A substantially pure polypeptideor protein will typically comprise about 50%, 60%, 70%, 80% or 90% W/Wof a protein sample, more usually about 95%, and preferably will be over99% pure. Protein purity or homogeneity may be indicated by a number ofmeans well-known in the art, such as polyacrylamide gel electrophoresisof a protein sample, followed by visualizing a single polypeptide bandupon staining the gel with a stain well-known in the art. For certainpurposes, higher resolution may be provided by using HPLC or other meanswell-known in the art for purification.

[0084] The term “polypeptide fragment” as used herein refers to apolypeptide of the instant invention that has an amino-terminal and/orcarboxy-terminal deletion compared to a full-length polypeptide. In apreferred embodiment, the polypeptide fragment is a contiguous sequencein which the amino acid sequence of the fragment is identical to thecorresponding positions in the naturally-occurring sequence. Fragmentstypically are at least 5, 6, 7, 8, 9 or 10 amino acids long, preferablyat least 12, 14, 16 or 18 amino acids long, more preferably at least 20amino acids long, more preferably at least 25, 30, 35, 40 or 45, aminoacids, even more preferably at least 50 or 60 amino acids long, and evenmore preferably at least 70 amino acids long.

[0085] A “derivative” refers to polypeptides or fragments thereof thatare substantially similar in primary structural sequence but whichinclude, e.g., in vivo or in vitro chemical and biochemicalmodifications that are not found in the native polypeptide. Suchmodifications include, for example, acetylation, acylation,ADP-ribosylation, amidation, covalent attachment of flavin, covalentattachment of a heme moiety, covalent attachment of a nucleotide ornucleotide derivative, covalent attachment of a lipid or lipidderivative, covalent attachment of phosphotidylinositol, cross-linking,cyclization, disulfide bond formation, demethylation, formation ofcovalent cross-links, formation of cystine, formation of pyroglutamate,formylation, gamma-carboxylation, glycosylation, GPI anchor formation,hydroxylation, iodination, methylation, myristoylation, oxidation,proteolytic processing, phosphorylation, prenylation, racemization,selenoylation, sulfation, transfer-RNA mediated addition of amino acidsto proteins such as arginylation, and ubiquitination. Other modificationinclude, e.g., labeling with radionuclides, and various enzymaticmodifications, as will be readily appreciated by those skilled in theart. A variety of methods for labeling polypeptides and of substituentsor labels useful for such purposes are well-known in the art, andinclude radioactive isotopes such as ¹²⁵I, ³²P, ³⁵S, and ³H, ligandswhich bind to labeled antiligands (e.g., antibodies), fluorophores,chcmiluminescent agents, enzymes, and antiligands which can serve asspecific binding pair members for a labeled ligand. The choice of labeldepends on the sensitivity required, ease of conjugation with theprimer, stability requirements, and available instrumentation. Methodsfor labeling polypeptides are well-known in the art. See Ausubel (1992),supra; Ausubel (1999), supra, herein incorporated by reference.

[0086] The term “fusion protein” refers to polypeptides of the instantinvention comprising polypeptides or fragments coupled to heterologousamino acid sequences. Fusion proteins are useful because they can beconstructed to contain two or more desired functional elements from twoor more different proteins. A fusion protein comprises at least 10contiguous amino acids from a polypeptide of interest, more preferablyat least 20 or 30 amino acids, even more preferably at least 40, 50 or60 amino acids, yet more preferably at least 75, 100 or 125 amino acids.Fusion proteins can be produced recombinantly by constructing a nucleicacid sequence which encodes the polypeptide or a fragment thereof inframe with a nucleic acid sequence encoding a different protein orpcptide and then expressing the fusion protein. Alternatively, a fusionprotein can be produced chemically by crosslinking the polypeptide or afragment thereof to another protein.

[0087] The term “analog” refers to both polypeptide analogs andnon-peptide analogs. The term “polypeptide analog” as used herein refersto a polypeptide of the instant invention that is comprised of a segmentof at least 25 amino acids that has substantial identity to a portion ofan amino acid sequence but which contains non-natural amino acids ornon-natural inter-residue bonds. In a preferred embodiment, the analoghas the same or similar biological activity as the native polypeptide.Typically, polypeptide analogs comprise a conservative amino acidsubstitution (or insertion or deletion) with respect to thenaturally-occurring sequence. Analogs typically are at least 20 aminoacids long, preferably at least 50 amino acids long or longer, and canoften be as long as a full-length naturally-occurring polypeptide.

[0088] The term “non-peptide analog” refers to a compound withproperties that are analogous to those of a reference polypeptide of theinstant invention. A non-peptide compound may also be termed a “peptidemimetic” or a “peptidomimetic.” Such compounds are often developed withthe aid of computerized molecular modeling. Peptide mimetics that arestructurally similar to useful peptides may be used to produce anequivalent effect. Generally, peptidomimetics are structurally similarto a paradigm polypeptide (i.e., a polypeptide that has a desiredbiochemical property or pharmacological activity), but have one or morepeptide linkages optionally replaced by a linkage selected from thegroup consisting of: ——CH₂NH——, ——CH₂S——, ——CH₂—CH₂——, ——CH═CH—— (cisand trans), ——COCH₂——, ——CH(OH)CH₂——, and —CH₂SO——, by methodswell-known in the art. Systematic substitution of one or more aminoacids of a consensus sequence with a D-amino acid of the same type(e.g., D-lysine in place of L-lysine) may also be used to generate morestable peptides. In addition, constrained peptides comprising aconsensus sequence or a substantially identical consensus sequencevariation may be generated by methods known in the art (Rizo et al.,Ann. Rev. Biochem. 61:387-418 (1992), incorporated herein by reference).For example, one may add internal cysteine residues capable of formingintramolecular disulfide bridges which cyclize the peptide.

[0089] A “polypeptide mutant” or “mutein” refers to a polypeptide of theinstant invention whose sequence contains substitutions, insertions ordeletions of one or more amino acids compared to the amino acid sequenceof a native or wild-type protein. A mutein may have one or more aminoacid point substitutions, in which a single amino acid at a position hasbeen changed to another amino acid, one or more insertions and/ordeletions, in which one or more amino acids are inserted or deleted,respectively, in the sequence of the naturally-occurring protein, and/ortruncations of the amino acid sequence at either or both the amino orcarboxy termini. Further, a mutein may have the same or differentbiological activity as the naturally-occurring protein. For instance, amutein may have an increased or decreased biological activity. A muteinhas at least 50% sequence similarity to the wild type protein, preferredis 60% sequence similarity, more preferred is 70% sequence similarity.Even more preferred are muteins having 80%, 85% or 90% sequencesimilarity to the wild type protein. In an even more preferredembodiment, a mutein exhibits 95% sequence identity, even morepreferably 97%, even more preferably 98% and even more preferably 99%.Sequence similarity may be measured by any common sequence analysisalgorithm, such as Gap or Bestfit.

[0090] Preferred amino acid substitutions are those which: (1) reducesusceptibility to proteolysis, (2) reduce susceptibility to oxidation,(3) alter binding affinity for forming protein complexes, (4) alterbinding affinity or enzymatic activity, and (5) confer or modify otherphysicochemical or functional properties of such analogs. For example,single or multiple amino acid substitutions (preferably conservativeamino acid substitutions) may be made in the naturally-occurringsequence (preferably in the portion of the polypeptide outside thedomain(s) forming intermolecular contacts. In a preferred embodiment,the amino acid substitutions are moderately conservative substitutionsor conservative substitutions. In a more preferred embodiment, the aminoacid substitutions are conservative substitutions. A conservative aminoacid substitution should not substantially change the structuralcharacteristics of the parent sequence (e.g., a replacement amino acidshould not tend to disrupt a helix that occurs in the parent sequence,or disrupt other types of secondary structure that characterizes theparent sequence). Examples of art-recognized polypeptide secondary andtertiary structures are described in Creighton (ed.), Proteins,Structures and Molecular Principles, W. H. Freeman and Company (1984);Branden et al. (ed.), Introduction to Protein Structure, GarlandPublishing (1991); Thornton et al., Nature 354:105-106 (1991), each ofwhich are incorporated herein by reference.

[0091] As used herein, the twenty conventional amino acids and theirabbreviations follow conventional usage. See Golub et al. (eds.),Immunology—A Synthesis 2^(nd) Ed., Sinauer Associates (1991), which isincorporated herein by reference. Stereoisomers (e.g., D-amino acids) ofthe twenty conventional amino acids, unnatural amino acids such as -,-disubstituted amino acids, N-alkyl amino acids, and otherunconventional amino acids may also be suitable components forpolypeptides of the present invention. Examples of unconventional aminoacids include: 4-hydroxyproline, γ-carboxyglutamate,-N,N,N-trimethyllysine, -N-acetyllysine, O-phosphoserine,N-acetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylysine,s-N-methylarginine, and other similar amino acids and imino acids (e.g.,4-hydroxyproline). In the polypeptide notation used herein, the lefthanddirection is the amino terminal direction and the right hand directionis the carboxy-terminal direction, in accordance with standard usage andconvention.

[0092] A protein has “homology” or is “homologous” to a protein fromanother organism if the encoded amino acid sequence of the protein has asimilar sequence to the encoded amino acid sequence of a protein of adifferent organism and has a similar biological activity or function.Alternatively, a protein may have homology or be homologous to anotherprotein if the two proteins have similar amino acid sequences and havesimilar biological activities or functions. Although two proteins aresaid to be “homologous,” this does not imply that there is necessarilyan evolutionary relationship between the proteins. Instead, the term“homologous” is defined to mean that the two proteins have similar aminoacid sequences and similar biological activities or functions. In apreferred embodiment, a homologous protein is one that exhibits 50%sequence similarity to the wild type protein, preferred is 60% sequencesimilarity, more preferred is 70% sequence similarity. Even morepreferred are homologous proteins that exhibit 80%, 85% or 90% sequencesimilarity to the wild type protein. In a yet more preferred embodiment,a homologous protein exhibits 95%, 97%, 98% or 99% sequence similarity.

[0093] When “sequence similarity” is used in reference to proteins orpeptides, it is recognized that residue positions that are not identicaloften differ by conservative amino acid substitutions. In a preferredembodiment, a polypeptide that has “sequence similarity” comprisesconservative or moderately conservative amino acid substitutions. A“conservative amino acid substitution” is one in which an amino acidresidue is substituted by another amino acid residue having a side chain(R group) with similar chemical properties (e.g., charge orhydrophobicity). In general, a conservative amino acid substitution willnot substantially change the functional properties of a protein. Incases where two or more amino acid sequences differ from each other byconservative substitutions, the percent sequence identity or degree ofsimilarity may be adjusted upwards to correct for the conservativenature of the substitution. Means for making this adjustment arewell-known to those of skill in the art. See, e.g., Pearson, MethodsMol. Biol. 24: 307-31 (1994), herein incorporated by reference.

[0094] For instance, the following six groups each contain amino acidsthat are conservative substitutions for one another: 1) Serine (S),Threonine (T); 2) Aspartic Acid (D), Glutamic Acid (E); 3) Asparagine(N), Glutamine (Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (I),Leucine (L), Methionine (M), Alanine (A), Valine (V), and 6)Phenylalanine (F), Tyrosine (Y), Tryptophan (W).

[0095] Alternatively, a conservative replacement is any change having apositive value in the PAM250 log-likelihood matrix disclosed in Gonnetet al., Science 256: 1443-45 (1992), herein incorporated by reference. A“moderately conservative” replacement is any change having a nonnegativevalue in the PAM250 log-likelihood matrix.

[0096] Sequence similarity for polypeptides, which is also referred toas sequence identity, is typically measured using sequence analysissoftware. Protein analysis software matches similar sequences usingmeasures of similarity assigned to various substitutions, deletions andother modifications, including conservative amino acid substitutions.For instance, GCG contains programs such as “Gap” and “Bestfit” whichcan be used with default parameters to determine sequence homology orsequence identity between closely related polypeptides, such ashomologous polypeptides from different species of organisms or between awild type protein and a mutein thereof. See, e.g., GCG Version 6.1.Other programs include FASTA, discussed supra.

[0097] A preferred algorithm when comparing a sequence of the inventionto a database containing a large number of sequences from differentorganisms is the computer program BLAST, especially blastp or tblastn.See, e.g., Altschul et al., J. Mol. Biol. 215: 403-410 (1990); Altschulet al., Nucleic Acids Res. 25:3389-402 (1997); herein incorporated byreference. Preferred parameters for blastp are: Expectation value: 10(default) Filter: seg (default) Cost to open a gap: 11 (default) Cost toextend a gap: 1 (default Max. alignments: 100 (default) Word size: 11(default) No. of descriptions: 100 (default) Penalty Matrix: BLOSUM62

[0098] The length of polypeptide sequences compared for homology willgenerally be at least about 16 amino acid residues, usually at leastabout 20 residues, more usually at least about 24 residues, typically atleast about 28 residues, and preferably more than about 35 residues.When searching a database containing sequences from a large number ofdifferent organisms, it is preferable to compare amino acid sequences.

[0099] Database searching using amino acid sequences can be measured byalgorithms other than blastp are known in the art. For instance,polypeptide sequences can be compared using FASTA, a program in GCGVersion 6.1. FASTA (e.g., FASTA2 and FASTA3) provides alignments andpercent sequence identity of the regions of the best overlap between thequery and search sequences (Pearson (1990), supra; Pearson (2000),supra. For example, percent sequence identity between amino acidsequences can be determined using FASTA with its default or recommendedparameters (a word size of 2 and the PAM250 scoring matrix), as providedin GCG Version 6.1, herein incorporated by reference.

[0100] An “antibody” refers to an intact immunoglobulin, or to anantigen-binding portion thereof that competes with the intact antibodyfor specific binding to a molecular species, e.g., a polypeptide of theinstant invention. Antigen-binding portions may be produced byrecombinant DNA techniques or by enzymatic or chemical cleavage ofintact antibodies. Antigen-binding portions include, inter alia, Fab,Fab′, F(ab′)₂, Fv, dAb, and complementarity determining region (CDR)fragments, single-chain antibodies (scFv), chimeric antibodies,diabodies and polypeptides that contain at least a portion of animmunoglobulin that is sufficient to confer specific antigen binding tothe polypeptide. An Fab fragment is a monovalent fragment consisting ofthe VL, VH, CL and CH1 domains; an F(ab′)₂ fragment is a bivalentfragment comprising two Fab fragments linked by a disulfide bridge atthe hinge region; an Fd fragment consists of the VH and CH1 domains; anFv fragment consists of the VL and VH domains of a single arm of anantibody; and a dAb fragment consists of a VH domain. See, e.g., Ward etal., Nature 341: 544-546 (1989).

[0101] By “bind specifically” and “specific binding” is here intendedthe ability of the antibody to bind to a first molecular species inpreference to binding to other molecular species with which the antibodyand first molecular species are admixed. An antibody is saidspecifically to “recognize” a first molecular species when it can bindspecifically to that first molecular species.

[0102] A single-chain antibody (scFv) is an antibody in which a VL andVH region are paired to form a monovalent molecule via a syntheticlinker that enables them to be made as a single protein chain. See,e.g., Bird et al., Science 242: 423-426 (1988); Huston et al., Proc.Natl. Acad. Sci. USA 85: 5879-5883 (1988). Diabodies are bivalent,bispecific antibodies in which VH and VL domains are expressed on asingle polypeptide chain, but using a linker that is too short to allowfor pairing between the two domains on the same chain, thereby forcingthe domains to pair with complementary domains of another chain andcreating two antigen binding sites. See e.g., Holliger et al., Proc.Natl. Acad. Sci. USA 90: 6444-6448 (1993); Poljak et al., Structure 2:1121-1123 (1994). One or more CDRs may be incorporated into a moleculeeither covalently or noncovalently to make it an immunoadhesin. Animmunoadhesin may incorporate the CDR(s) as part of a larger polypeptidechain, may covalently link the CDR(s) to another polypeptide chain, ormay incorporate the CDR(s) noncovalently. The CDRs permit theimmunoadhesin to specifically bind to a particular antigen of interest.A chimeric antibody is an antibody that contains one or more regionsfrom one antibody and one or more regions from one or more otherantibodies.

[0103] An antibody may have one or more binding sites. If there is morethan one binding site, the binding sites may be identical to one anotheror may be different. For instance, a naturally-occurring immunoglobulinhas two identical binding sites, a single-chain antibody or Fab fragmenthas one binding site, while a “bispecific” or “bifunctional” antibodyhas two different binding sites.

[0104] An “isolated antibody” is an antibody that (1) is not associatedwith naturally-associated components, including othernaturally-associated antibodies, that accompany it in its native state,(2) is free of other proteins from the same species, (3) is expressed bya cell from a different species, or (4) does not occur in nature. It isknown that purified proteins, including purified antibodies, may bestabilized with non-naturally-associated components. Thenon-naturally-associated component may be a protein, such as albumin(e.g., BSA) or a chemical such as polyethylene glycol (PEG).

[0105] A “neutralizing antibody” or “an inhibitory antibody” is anantibody that inhibits the activity of a polypeptide or blocks thebinding of a polypeptide to a ligand that normally binds to it. An“activating antibody” is an antibody that increases the activity of apolypeptide.

[0106] The term “epitope” includes any protein determinant capable ofspecifically binding to an immunoglobulin or T-cell receptor. Epitopicdeterminants usually consist of chemically active surface groupings ofmolecules such as amino acids or sugar side chains and usually havespecific three-dimensional structural characteristics, as well asspecific charge characteristics. An antibody is said to specificallybind an antigen when the dissociation constant is less than 1 μM,preferably less than 100 nM and most preferably less than 10 nM.

[0107] The term “patient” as used herein includes human and veterinarysubjects.

[0108] Throughout this specification and claims, the word “comprise,” orvariations such as “comprises” or “comprising,” will be understood toimply the inclusion of a stated integer or group of integers but not theexclusion of any other integer or group of integers.

[0109] The term “breast specific” refers to a nucleic acid molecule orpolypeptide that is expressed predominantly in the breast as compared toother tissues in the body. In a preferred embodiment, a “breastspecific” nucleic acid molecule or polypeptide is expressed at a levelthat is 5-fold higher than any other tissue in the body. In a morepreferred embodiment, the “breast specific” nucleic acid molecule orpolypeptide is expressed at a level that is 10-fold higher than anyother tissue in the body, more preferably at least 15-fold, 20-fold,25-fold, 50-fold or 100-fold higher than any other tissue in the body.Nucleic acid molecule levels may be measured by nucleic acidhybridization, such as Northern blot hybridization, or quantitative PCR.Polypeptide levels may be measured by any method known to accuratelyquantitate protein levels, such as Western blot analysis.

[0110] Nucleic Acid Molecules, Regulatory Sequences, Vectors, Host Cellsand Recombinant Methods of Making Polypeptides

[0111] Nucleic Acid Molecules

[0112] One aspect of the invention provides isolated nucleic acidmolecules that are specific to the breast or to breast cells or tissueor that are derived from such nucleic acid molecules. These isolatedbreast specific nucleic acids (BSNAs) may comprise a cDNA, a genomicDNA, RNA, or a fragment of one of these nucleic acids, or may be anon-naturally-occurring nucleic acid molecule. In a preferredembodiment, the nucleic acid molecule encodes a polypeptide that isspecific to breast, a breast-specific polypeptide (BSP). In a morepreferred embodiment, the nucleic acid molecule encodes a polypeptidethat comprises an amino acid sequence of SEQ ID NO: 160 through 282. Inanother highly preferred embodiment, the nucleic acid molecule comprisesa nucleic acid sequence of SEQ ID NO: 1 through 159.

[0113] A BSNA may be derived from a human or from another animal. In apreferred embodiment, the BSNA is derived from a human or other mammal.In a more preferred embodiment, the BSNA is derived from a human orother primate. In an even more preferred embodiment, the BSNA is derivedfrom a human.

[0114] By “nucleic acid molecule” for purposes of the present invention,it is also meant to be inclusive of nucleic acid sequences thatselectively hybridize to a nucleic acid molecule encoding a BSNA or acomplement thereof. The hybridizing nucleic acid molecule may or may notencode a polypeptide or may not encode a BSP. However, in a preferredembodiment, the hybridizing nucleic acid molecule encodes a BSP. In amore preferred embodiment, the invention provides a nucleic acidmolecule that selectively hybridizes to a nucleic acid molecule thatencodes a polypeptide comprising an amino acid sequence of SEQ ID NO:160 through 282. In an even more preferred embodiment, the inventionprovides a nucleic acid molecule that selectively hybridizes to anucleic acid molecule comprising the nucleic acid sequence of SEQ ID NO:1 through 159.

[0115] In a preferred embodiment, the nucleic acid molecule selectivelyhybridizes to a nucleic acid molecule encoding a BSP under lowstringency conditions. In a more preferred embodiment, the nucleic acidmolecule selectively hybridizes to a nucleic acid molecule encoding aBSP under moderate stringency conditions. In a more preferredembodiment, the nucleic acid molecule selectively hybridizes to anucleic acid molecule encoding a BSP under high stringency conditions.In an even more preferred embodiment, the nucleic acid moleculehybridizes under low, moderate or high stringency conditions to anucleic acid molecule encoding a polypeptide comprising an amino acidsequence of SEQ ID NO: 160 through 282. In a yet more preferredembodiment, the nucleic acid molecule hybridizes under low, moderate orhigh stringency conditions to a nucleic acid molecule comprising anucleic acid sequence selected from SEQ ID NO: 1 through 159. In apreferred embodiment of the invention, the hybridizing nucleic acidmolecule may be used to express recombinantly a polypeptide of theinvention.

[0116] By “nucleic acid molecule” as used herein it is also meant to beinclusive of sequences that exhibits substantial sequence similarity toa nucleic acid encoding a BSP or a complement of the encoding nucleicacid molecule. In a preferred embodiment, the nucleic acid moleculeexhibits substantial sequence similarity to a nucleic acid moleculeencoding human BSP. In a more preferred embodiment, the nucleic acidmolecule exhibits substantial sequence similarity to a nucleic acidmolecule encoding a polypeptide having an amino acid sequence of SEQ IDNO: 160 through 282. In a preferred embodiment, the similar nucleic acidmolecule is one that has at least 60% sequence identity with a nucleicacid molecule encoding a BSP, such as a polypeptide having an amino acidsequence of SEQ ID NO: 160 through 282, more preferably at least 70%,even more preferably at least 80% and even more preferably at least 85%.In a more preferred embodiment, the similar nucleic acid molecule is onethat has at least 90% sequence identity with a nucleic acid moleculeencoding a BSP, more preferably at least 95%, more preferably at least97%, even more preferably at least 98%, and still more preferably atleast 99%. In another highly preferred embodiment, the nucleic acidmolecule is one that has at least 99.5%, 99.6%, 99.7%, 99.8% or 99.9%sequence identity with a nucleic acid molecule encoding a BSP.

[0117] In another preferred embodiment, the nucleic acid moleculeexhibits substantial sequence similarity to a BSNA or its complement. Ina more preferred embodiment, the nucleic acid molecule exhibitssubstantial sequence similarity to a nucleic acid molecule comprising anucleic acid sequence of SEQ ID NO: 1 through 159. In a preferredembodiment, the nucleic acid molecule is one that has at least 60%sequence identity with a BSNA, such as one having a nucleic acidsequence of SEQ ID NO: 1 through 159, more preferably at least 70%, evenmore preferably at least 80% and even more preferably at least 85%. In amore preferred embodiment, the nucleic acid molecule is one that has atleast 90% sequence identity with a BSNA, more preferably at least 95%,more preferably at least 97%, even more preferably at least 98%, andstill more preferably at least 99%. In another highly preferredembodiment, the nucleic acid molecule is one that has at least 99.5%,99.6%, 99.7%, 99.8% or 99.9% sequence identity with a BSNA.

[0118] A nucleic acid molecule that exhibits substantial sequencesimilarity may be one that exhibits sequence identity over its entirelength to a BSNA or to a nucleic acid molecule encoding a BSP, or may beone that is similar over only a part of its length. In this case, thepart is at least 50 nucleotides of the BSNA or the nucleic acid moleculeencoding a BSP, preferably at least 100 nucleotides, more preferably atleast 150 or 200 nucleotides, even more preferably at least 250 or 300nucleotides, still more preferably at least 400 or 500 nucleotides.

[0119] The substantially similar nucleic acid molecule may be anaturally-occurring one that is derived from another species, especiallyone derived from another primate, wherein the similar nucleic acidmolecule encodes an amino acid sequence that exhibits significantsequence identity to that of SEQ ID NO: 160 through 282 or demonstratessignificant sequence identity to the nucleotide sequence of SEQ ID NO: 1through 159. The similar nucleic acid molecule may also be anaturally-occurring nucleic acid molecule from a human, when the BSNA isa member of a gene family. The similar nucleic acid molecule may also bea naturally-occurring nucleic acid molecule derived from a non-primate,mammalian species, including without limitation, domesticated species,e.g., dog, cat, mouse, rat, rabbit, hamster, cow, horse and pig; andwild animals, e.g., monkey, fox, lions, tigers, bears, giraffes, zebras,etc. The substantially similar nucleic acid molecule may also be anaturally-occurring nucleic acid molecule derived from a non-mammalianspecies, such as birds or reptiles. The naturally-occurringsubstantially similar nucleic acid molecule may be isolated directlyfrom humans or other species. In another embodiment, the substantiallysimilar nucleic acid molecule may be one that is experimentally producedby random mutation of a nucleic acid molecule. In another embodiment,the substantially similar nucleic acid molecule may be one that isexperimentally produced by directed mutation of a BSNA. Further, thesubstantially similar nucleic acid molecule may or may not be a BSNA.However, in a preferred embodiment, the substantially similar nucleicacid molecule is a BSNA.

[0120] By “nucleic acid molecule” it is also meant to be inclusive ofallelic variants of a BSNA or a nucleic acid encoding a BSP. Forinstance, single nucleotide polymorphisms (SNPs) occur frequently ineukaryotic genomes. In fact, more than 1.4 million SNPs have alreadyidentified in the human genome, International Human Genome SequencingConsortium, Nature 409: 860-921 (2001). Thus, the sequence determinedfrom one individual of a species may differ from other allelic formspresent within the population. Additionally, small deletions andinsertions, rather than single nucleotide polymorphisms, are notuncommon in the general population, and often do not alter the functionof the protein. Further, amino acid substitutions occur frequently amongnatural allelic variants, and often do not substantially change proteinfunction.

[0121] In a preferred embodiment, the nucleic acid molecule comprisingan allelic variant is a variant of a gene, wherein the gene istranscribed into an mRNA that encodes a BSP. In a more preferredembodiment, the gene is transcribed into an mRNA that encodes a BSPcomprising an amino acid sequence of SEQ ID NO: 160 through 282. Inanother preferred embodiment, the allelic variant is a variant of agene, wherein the gene is transcribed into an mRNA that is a BSNA. In amore preferred embodiment, the gene is transcribed into an mRNA thatcomprises the nucleic acid sequence of SEQ ID NO: 1 through 159. In apreferred embodiment, the allelic variant is a naturally-occurringallelic variant in the species of interest. In a more preferredembodiment, the species of interest is human.

[0122] By “nucleic acid molecule” it is also meant to be inclusive of apart of a nucleic acid sequence of the instant invention. The part mayor may not encode a polypeptide, and may or may not encode a polypeptidethat is a BSP. However, in a preferred embodiment, the part encodes aBSP. In one aspect, the invention comprises a part of a BSNA. In asecond aspect, the invention comprises a part of a nucleic acid moleculethat hybridizes or exhibits substantial sequence similarity to a BSNA.In a third aspect, the invention comprises a part of a nucleic acidmolecule that is an allelic variant of a BSNA. In a fourth aspect, theinvention comprises a part of a nucleic acid molecule that encodes aBSP. A part comprises at least 10 nucleotides, more preferably at least15, 17, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250,300, 350, 400 or 500 nucleotides. The maximum size of a nucleic acidpart is one nucleotide shorter than the sequence of the nucleic acidmolecule encoding the full-length protein.

[0123] By “nucleic acid molecule” it is also meant to be inclusive ofsequence that encoding a fusion protein, a homologous protein, apolypeptide fragment, a mutein or a polypeptide analog, as describedbelow.

[0124] Nucleotide sequences of the instantly-described nucleic acidswere determined by sequencing a DNA molecule that had resulted, directlyor indirectly, from at least one enzymatic polymerization reaction(e.g., reverse transcription and/or polymerase chain reaction) using anautomated sequencer (such as the MegaBACE™ 1000, Molecular Dynamics,Sunnyvale, Calif., USA). Further, all amino acid sequences of thepolypeptides of the present invention were predicted by translation fromthe nucleic acid sequences so determined, unless otherwise specified.

[0125] In a preferred embodiment of the invention, the nucleic acidmolecule contains modifications of the native nucleic acid molecule.These modifications include nonnative internucleoside bonds,post-synthetic modifications or altered nucleotide analogues. One havingordinary skill in the art would recognize that the type of modificationthat can be made will depend upon the intended use of the nucleic acidmolecule. For instance, when the nucleic acid molecule is used as ahybridization probe, the range of such modifications will be limited tothose that permit sequence-discriminating base pairing of the resultingnucleic acid. When used to direct expression of RNA or protein in vitroor in vivo, the range of such modifications will be limited to thosethat permit the nucleic acid to function properly as a polymerizationsubstrate. When the isolated nucleic acid is used as a therapeuticagent, the modifications will be limited to those that do not confertoxicity upon the isolated nucleic acid.

[0126] In a preferred embodiment, isolated nucleic acid molecules caninclude nucleotide analogues that incorporate labels that are directlydetectable, such as radiolabels or fluorophores, or nucleotide analoguesthat incorporate labels that can be visualized in a subsequent reaction,such as biotin or various haptens. In a more preferred embodiment, thelabeled nucleic acid molecule may be used as a hybridization probe.

[0127] Common radiolabeled analogues include those labeled with ³³P,³²P, and 35S, such as -³²P-dATP, -³²P-dCTP, -³²P-dGTP, -³²P-dTTP,-³²P-3′dATP, -³²P-ATP, -³²P-CTP, -³²P-GTP, -³²P-UTP, -³⁵S-dATP,α-³⁵S-GTP, α-³³P-dATP, and the like.

[0128] Commercially available fluorescent nucleotide analogues readilyincorporated into the nucleic acids of the present invention includeCy3-dCTP, Cy3-dUTP, Cy5-dCTP, Cy3-dUTP (Amersham Pharmacia Biotech,Piscataway, N.J., USA), fluorescein-12-dUTP,tetramethylrhodamine-6-dUTP, Texas Red®-5-dUTP, Cascade Blue®-7-dUTP,BODIPY® FL-14-dUTP, BODIPY® TMR-14-dUTP, BODIPY® TR-14-dUTP, RhodamineGreen™-5-dUTP, Oregon Green® 488-5-dUTP, Texas Red®-12-dUTP, BODIPY®630/650-14-dUTP, BODIPY® 650/665-14-dUTP, Alexa Fluor® 488-5-dUTP, AlexaFluor® 532-5-dUTP, Alexa Fluor® 568-5-dUTP, Alexa Fluor® 594-5-dUTP,Alexa Fluor® 546-14-dUTP, fluorescein-12-UTP,tetramethylrhodamine-6-UTP, Texas Red®-5-UTP, Cascade Blue®-7-UTP,BODIPY® FL-14-UTP, BODIPY® TMR-14-UTP, BODIPY® TR-14-UTP, RhodamineGreen™-5-UTP, Alexa Fluor® 488-5-UTP, Alexa Fluor® 546-14-UTP (MolecularProbes, Inc. Eugene, Oreg., USA). One may also custom synthesizenucleotides having other fluorophores. See Henegariu et al., NatureBiotechnol. 18: 345-348 (2000), the disclosure of which is incorporatedherein by reference in its entirety.

[0129] Haptens that are commonly conjugated to nucleotides forsubsequent labeling include biotin (biotin-11-dUTP, Molecular Probes,Inc., Eugene, Oreg., USA; biotin-21-UTP, biotin-21-dUTP, ClontechLaboratories, Inc., Palo Alto, Calif., USA), digoxigenin (DIG-11-dUTP,alkali labile, DIG-11-UTP, Roche Diagnostics Corp., Indianapolis, Ind.,USA), and dinitrophenyl (dinitrophenyl-11-dUTP, Molecular Probes, Inc.,Eugene, Oreg., USA).

[0130] Nucleic acid molecules can be labeled by incorporation of labelednucleotide analogues into the nucleic acid. Such analogues can beincorporated by enzymatic polymerization, such as by nick translation,random priming, polymerase chain reaction (PCR), terminal transferasetailing, and end-filling of overhangs, for DNA molecules, and in vitrotranscription driven, e.g., from phage promoters, such as T7, T3, andSP6, for RNA molecules. Commercial kits are readily available for eachsuch labeling approach. Analogues can also be incorporated duringautomated solid phase chemical synthesis. Labels can also beincorporated after nucleic acid synthesis, with the 5′ phosphate and 3′hydroxyl providing convenient sites for post-synthetic covalentattachment of detectable labels.

[0131] Other post-synthetic approaches also permit internal labeling ofnucleic acids. For example, fluorophores can be attached using acisplatin reagent that reacts with the N7 of guanine residues (and, to alesser extent, adenine bases) in DNA, RNA, and PNA to provide a stablecoordination complex between the nucleic acid and fluorophore label(Universal Linkage System) (available from Molecular Probes, Inc.,Eugene, Oreg., USA and Amersham Pharmacia Biotech, Piscataway, N.J.,USA); see Alers et al., Genes, Chromosomes & Cancer 25: 301-305 (1999);Jelsma et al., J. NIH Res. 5: 82 (1994); Van Belkum et al.,BioTechniques 16: 148-153 (1994), incorporated herein by reference. Asanother example, nucleic acids can be labeled using adisulfide-containing linker (FastTag™ Reagent, Vector Laboratories,Inc., Burlingame, Calif., USA) that is photo- or thermally-coupled tothe target nucleic acid using aryl azide chemistry; after reduction, afree thiol is available for coupling to a hapten, fluorophore, sugar,affinity ligand, or other marker.

[0132] One or more independent or interacting labels can be incorporatedinto the nucleic acid molecules of the present invention. For example,both a fluorophore and a moiety that in proximity thereto acts to quenchfluorescence can be included to report specific hybridization throughrelease of fluorescence quenching or to report exonucleotidic excision.See, e.g., Tyagi et al., Nature Biotechnol. 14: 303-308 (1996); Tyagi etal., Nature Biotechnol. 16: 49-53 (1998); Sokol et al., Proc. Natl.Acad. Sci. USA 95: 11538-11543 (1998); Kostrikis et al., Science 279:1228-1229 (1998); Marras et al., Genet. Anal. 14: 151-156 (1999); U.S.Pat. Nos. 5,846,726; 5,925,517; 5,925,517; 5,723,591 and 5,538,848;Holland et al., Proc. Natl. Acad. Sci. USA 88: 7276-7280 (1991); Heid etal., Genome Res. 6(10): 986-94 (1996); Kuimelis et al., Nucleic AcidsSymp. Ser. (37): 255-6 (1997); the disclosures of which are incorporatedherein by reference in their entireties.

[0133] Nucleic acid molecules of the invention may be modified byaltering one or more native phosphodiester internucleoside bonds to morenuclease-resistant, internucleoside bonds. See Hartmann et al. (eds.),Manual of Antisense Methodology: Perspectives in Antisense Science,Kluwer Law International (1999); Stein et al. (eds.), Applied AntisenseOligonucleotide Technology, Wiley-Liss (1998); Chadwick et al. (eds.),Oligonucleotides as Therapeutic Agents—Symposium No. 209, John Wiley &Son Ltd (1997); the disclosures of which are incorporated herein byreference in their entireties. Such altered internucleoside bonds areoften desired for antisense techniques or for targeted gene correction.See Gamper et al., Nucl. Acids Res. 28(21): 4332-4339 (2000), thedisclosure of which is incorporated herein by reference in its entirety.

[0134] Modified oligonucleotide backbones include, without limitation,phosphorothioates, chiral phosphorothioates, phosphorodithioates,phosphotriesters, aminoalkylphosphotriesters, methyl and other alkylphosphonates including 3′-alkylene phosphonates and chiral phosphonates,phosphinates, phosphoramidates including 3′-amino phosphoramidate andaminoalkylphosphoramidates, thionophosphoramidates,thionoalkylphosphonates, thionoalkylphosphotriesters, andboranophosphates having normal 3′-5′ linkages, 2′-5′ linked analogs ofthese, and those having inverted polarity wherein the adjacent pairs ofnucleoside units are linked 3′-5′ to 5′-3′ or 2′-5′ to 5′-2′.Representative United States patents that teach the preparation of theabove phosphorus-containing linkages include, but are not limited to,U.S. Pat. Nos. 3,687,808; 4,469,863; 4,476,301; 5,023,243; 5,177,196;5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,717; 5,321,131;5,399,676; 5,405,939; 5,453,496; 5,455,233; 5,466,677; 5,476,925;5,519,126; 5,536,821; 5,541,306; 5,550,111; 5,563,253; 5,571,799;5,587,361; and 5,625,050, the disclosures of which are incorporatedherein by reference in their entireties. In a preferred embodiment, themodified internucleoside linkages may be used for antisense techniques.

[0135] Other modified oligonucleotide backbones do not include aphosphorus atom, but have backbones that are formed by short chain alkylor cycloalkyl internucleoside linkages, mixed heteroatom and alkyl orcycloalkyl internucleoside linkages, or one or more short chainheteroatomic or heterocyclic internucleoside linkages. These includethose having morpholino linkages (formed in part from the sugar portionof a nucleoside); siloxane backbones; sulfide, sulfoxide and sulfonebackbones; formacetyl and thioformacetyl backbones; methylene formacetyland thioformacetyl backbones; alkene containing backbones; sulfamatebackbones; methyleneimino and methylenehydrazino backbones; sulfonateand sulfonamide backbones; amide backbones; and others having mixed N,O, S and CH₂ component parts. Representative U.S. patents that teach thepreparation of the above backbones include, but are not limited to, U.S.Pat. Nos. 5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141;5,235,033; 5,264,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677;5,470,967; 5,489,677; 5,541,307; 5,561,225; 5,596,086; 5,602,240;5,610,289; 5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070;5,663,312; 5,633,360; 5,677,437 and 5,677,439; the disclosures of whichare incorporated herein by reference in their entireties.

[0136] In other preferred oligonucleotide mimetics, both the sugar andthe internucleoside linkage are replaced with novel groups, such aspeptide nucleic acids (PNA). In PNA compounds, the phosphodiesterbackbone of the nucleic acid is replaced with an amide-containingbackbone, in particular by repeating N-(2-aminoethyl) glycine unitslinked by amide bonds. Nucleobases are bound directly or indirectly toaza nitrogen atoms of the amide portion of the backbone, typically bymethylene carbonyl linkages. PNA can be synthesized using a modifiedpeptide synthesis protocol. PNA oligomers can be synthesized by bothFmoc and tBoc methods. Representative U.S. patents that teach thepreparation of PNA compounds include, but are not limited to, U.S Pat.Nos. 5,539,082; 5,714,331; and 5,719,262, each of which is hereinincorporated by reference. Automated PNA synthesis is readily achievableon commercial synthesizers (see, e.g., “PNA User's Guide,” Rev. 2,February 1998, Perseptive Biosystems Part No. 60138, Applied Biosystems,Inc., Foster City, Calif.).

[0137] PNA molecules are advantageous for a number of reasons. First,because the PNA backbone is uncharged, PNA/DNA and PNA/RNA duplexes havea higher thermal stability than is found in DNA/DNA and DNA/RNAduplexes. The T_(m) of a PNA/DNA or PNA/RNA duplex is generally 1° C.higher per base pair than the Tm of the corresponding DNA/DNA or DNA/RNAduplex (in 100 mM NaCl). Second, PNA molecules can also form stablePNA/DNA complexes at low ionic strength, under conditions in whichDNA/DNA duplex formation does not occur. Third, PNA also demonstratesgreater specificity in binding to complementary DNA because a PNA/DNAmismatch is more destabilizing than DNA/DNA mismatch. A single mismatchin mixed a PNA/DNA 15-mer lowers the Tm by 8-20° C. (15° C. on average).In the corresponding DNA/DNA duplexes, a single mismatch lowers the Tmby 4-16° C. (11° C. on average). Because PNA probes can be significantlyshorter than DNA probes, their specificity is greater. Fourth, PNAoligomers are resistant to degradation by enzymes, and the lifetime ofthese compounds is extended both in vivo and in vitro because nucleasesand proteases do not recognize the PNA polyamide backbone withnucleobase sidechains. See, e.g., Ray et al., FASEB J. 14(9): 1041-60(2000); Nielsen et al., Pharmacol Toxicol. 86(1): 3-7 (2000); Larsen etal., Biochim Biophys Acta. 1489(1): 159-66 (1999); Nielsen, Curr. Opin.Struct. Biol. 9(3): 353-7 (1999), and Nielsen, Curr. Opin. Biotechnol.10(1): 71-5 (1999), the disclosures of which are incorporated herein byreference in their entireties.

[0138] Nucleic acid molecules may be modified compared to their nativestructure throughout the length of the nucleic acid molecule or can belocalized to discrete portions thereof. As an example of the latter,chimeric nucleic acids can be synthesized that have discrete DNA and RNAdomains and that can be used for targeted gene repair and modified PCRreactions, as further described in U.S. Pat. Nos. 5,760,012 and5,731,181, Misra et al., Biochem. 37: 1917-1925 (1998); and Finn et al.,Nucl. Acids Res. 24: 3357-3363 (1996), the disclosures of which areincorporated herein by reference in their entireties.

[0139] Unless otherwise specified, nucleic acids of the presentinvention can include any topological conformation appropriate to thedesired use; the term thus explicitly comprehends, among others,single-stranded, double-stranded, triplexed, quadruplexed, partiallydouble-stranded, partially-triplexed, partially-quadruplexed, branched,hairpinned, circular, and padlocked conformations. Padlock conformationsand their utilities are further described in Baner et al., Curr. Opin.Biotechnol. 12: 11-15 (2001); Escude et al., Proc. Natl. Acad. Sci. USA14: 96(19):10603-7 (1999); Nilsson et al., Science 265(5181): 2085-8(1994), the disclosures of which are incorporated herein by reference intheir entireties. Triplex and quadruplex conformations, and theirutilities, are reviewed in Praseuth et al., Biochim. Biophys. Acta.1489(1): 181-206 (1999); Fox, Curr. Med. Chem. 7(1): 17-37 (2000);Kochetkova et al., Methods Mol. Biol. 130: 189-201 (2000); Chan et al.,J. Mol. Med. 75(4): 267-82 (1997), the disclosures of which areincorporated herein by reference in their entireties.

[0140] Methods for Using Nucleic Acid Molecules as Probes and Primers

[0141] Thc isolated nucleic acid molecules of the present invention canbe used as hybridization probes to detect, characterize, and quantifyhybridizing nucleic acids in, and isolate hybridizing nucleic acidsfrom, both genomic and transcript-derived nucleic acid samples. Whenfree in solution, such probes are typically, but not invariably,detectably labeled; bound to a substrate, as in a microarray, suchprobes are typically, but not invariably unlabeled.

[0142] In one embodiment, the isolated nucleic acids of the presentinvention can be used as probes to detect and characterize grossalterations in the gene of a BSNA, such as deletions, insertions,translocations, and duplications of the BSNA genomic locus throughfluorescence in situ hybridization (FISH) to chromosome spreads. See,e.g., Andreeff et al. (eds.), Introduction to Fluorescence In SituHybridization: Principles and Clinical Applications, John Wiley & Sons(1999), the disclosure of which is incorporated herein by reference inits entirety. The isolated nucleic acids of the present invention can beused as probes to assess smaller genomic alterations using, e.g.,Southern blot detection of restriction fragment length polymorphisms.The isolated nucleic acid molecules of the present invention can be usedas probes to isolate genomic clones that include the nucleic acidmolecules of the present invention, which thereafter can be restrictionmappcd and sequenced to identify deletions, insertions, translocations,and substitutions (single nucleotide polymorphisms, SNPs) at thesequence level.

[0143] In another embodiment, the isolated nucleic acid molecules of thepresent invention can be used as probes to detect, characterize, andquantify BSNA in, and isolate BSNA from, transcript-derived nucleic acidsamples. In one aspect, the isolated nucleic acid molecules of thepresent invention can be used as hybridization probes to detect,characterize by length, and quantify mRNA by Northern blot of total orpoly-A⁺-selected RNA samples. In another aspect, the isolated nucleicacid molecules of the present invention can be used as hybridizationprobes to detect, characterize by location, and quantify mRNA by in situhybridization to tissue sections. See, e.g., Schwarchzacher et al., InSitu Hybridization, Springer-Verlag New York (2000), the disclosure ofwhich is incorporated herein by reference in its entirety. In anotherpreferred embodiment, the isolated nucleic acid molecules of the presentinvention can be used as hybridization probes to measure therepresentation of clones in a cDNA library or to isolate hybridizingnucleic acid molecules acids from cDNA libraries, permitting sequencelevel characterization of mRNAs that hybridize to BSNAs, including,without limitations, identification of deletions, insertions,substitutions, truncations, alternatively spliced forms and singlenucleotide polymorphisms. In yet another preferred embodiment, thenucleic acid molecules of the instant invention may be used inmicroarrays.

[0144] All of the aforementioned probe techniques are well within theskill in the art, and are described at greater length in standard textssuch as Sambrook (2001), supra; Ausubel (1999), supra; and Walker et al.(eds.), The Nucleic Acids Protocols Handbook, Humana Press (2000), thedisclosures of which are incorporated herein by reference in theirentirety.

[0145] Thus, in one embodiment, a nucleic acid molecule of the inventionmay be used as a probe or primer to identify or amplify a second nucleicacid molecule that selectively hybridizes to the nucleic acid moleculeof the invention. In a preferred embodiment, the probe or primer isderived from a nucleic acid molecule encoding a BSP. In a more preferredembodiment, the probe or primer is derived from a nucleic acid moleculeencoding a polypeptide having an amino acid sequence of SEQ ID NO: 160through 282. In another preferred embodiment, the probe or primer isderived from a BSNA. In a more preferred embodiment, the probe or primeris derived from a nucleic acid molecule having a nucleotide sequence ofSEQ ID NO: 1 through 159.

[0146] In general, a probe or primer is at least 10 nucleotides inlength, more preferably at least 12, more preferably at least 14 andeven more preferably at least 16 or 17 nucleotides in length. In an evenmore preferred embodiment, the probe or primer is at least 18nucleotides in length, even more preferably at least 20 nucleotides andeven more preferably at least 22 nucleotides in length. Primers andprobes may also be longer in length. For instance, a probe or primer maybe 25 nucleotides in length, or may be 30, 40 or 50 nucleotides inlength. Methods of performing nucleic acid hybridization usingoligonucleotide probes are well-known in the art. See, e.g., Sambrook etal., 1989, supra, Chapter 11 and pp. 11.31-11.32 and 11.40-11.44, whichdescribes radiolabeling of short probes, and pp. 11.45-11.53, whichdescribe hybridization conditions for oligonucleotide probes, includingspecific conditions for probe hybridization (pp. 11.50-11.51).

[0147] Methods of performing primer-directed amplification are alsowell-known in the art. Methods for performing the polymerase chainreaction (PCR) are compiled, inter alia, in McPherson, PCR Basics: FromBackground to Bench, Springer Verlag (2000); Innis et al. (eds.), PCRApplications: Protocols for Functional Genomics, Academic Press (1999);Gelfand et al. (eds.), PCR Strategies, Academic Press (1998); Newton etal., PCR, Springer-Verlag New York (1997); Burke (ed.), PCR: EssentialTechniques, John Wiley & Son Ltd (1996); White (ed.), PCR CloningProtocols: From Molecular Cloning to Genetic Engineering, Vol. 67,Humana Press (1996); McPherson et al. (eds.), PCR 2: A PracticalApproach, Oxford University Press, Inc. (1995); the disclosures of whichare incorporated herein by reference in their entireties. Methods forperforming RT-PCR are collected, e.g., in Siebert et al. (eds.), GeneCloning and Analysis by RT-PCR, Eaton Publishing Company/Bio TechniquesBooks Division, 1998; Siebert (ed.), PCR Technique:RT-PCR, EatonPublishing Company/BioTechniques Books (1995); the disclosure of whichis incorporated herein by reference in its entirety.

[0148] PCR and hybridization methods may be used to identify and/orisolate allelic variants, homologous nucleic acid molecules andfragments of the nucleic acid molecules of the invention. PCR andhybridization methods may also be used to identify, amplify and/orisolate nucleic acid molecules that encode homologous proteins, analogs,fusion protein or muteins of the invention. The nucleic acid primers ofthe present invention can be used to prime amplification of nucleic acidmolecules of the invention, using transcript-derived or genomic DNA astemplate.

[0149] The nucleic acid primers of the present invention can also beused, for example, to prime single base extension (SBE) for SNPdetection (See, e.g., U.S. Pat. No. 6,004,744, the disclosure of whichis incorporated herein by reference in its entirety).

[0150] Isothermal amplification approaches, such as rolling circleamplification, are also now well-described. See, e.g., Schweitzer etal., Curr. Opin. Biotechnol. 12(1): 21-7 (2001); U.S. Pat. Nos.5,854,033 and 5,714,320; and international patent publications WO97/19193 and WO 00/15779, the disclosures of which are incorporatedherein by reference in their entireties. Rolling circle amplificationcan be combined with other techniques to facilitate SNP detection. See,e.g., Lizardi et al., Nature Genet. 19(3): 225-32 (1998).

[0151] Nucleic acid molecules of the present invention may be bound to asubstrate either covalently or noncovalently. The substrate can beporous or solid, planar or non-planar, unitary or distributed. The boundnucleic acid molecules may be used as hybridization probes, and may belabeled or unlabeled. In a preferred embodiment, the bound nucleic acidmolecules are unlabeled.

[0152] In one embodiment, the nucleic acid molecule of the presentinvention is bound to a porous substrate, e.g., a membrane, typicallycomprising nitrocellulose, nylon, or positively-charged derivatizednylon. The nucleic acid molecule of the present invention can be used todetect a hybridizing nucleic acid molecule that is present within alabeled nucleic acid sample, e.g., a sample of transcript-derivednucleic acids. In another embodiment, the nucleic acid molecule is boundto a solid substrate, including, without limitation, glass, amorphoussilicon, crystalline silicon or plastics. Examples of plastics include,without limitation, polymethylacrylic, polyethylene, polypropylene,polyacrylate, polymethylmethacrylate, polyvinylchloride,polytetrafluoroethylene, polystyrene, polycarbonate, polyacetal,polysulfone, celluloseacetate, cellulosenitrate, nitrocellulose, ormixtures thereof The solid substrate may be any shape, includingrectangular, disk-like and spherical. In a preferred embodiment, thesolid substrate is a microscope slide or slide-shaped substrate.

[0153] The nucleic acid molecule of the present invention can beattached covalently to a surface of the support substrate or applied toa derivatized surface in a chaotropic agent that facilitatesdenaturation and adherence by presumed noncovalent interactions, or somecombination thcrcof. The nucleic acid molecule of the present inventioncan be bound to a substrate to which a plurality of other nucleic acidsare concurrently bound, hybridization to each of the plurality of boundnucleic acids being separately detectable. At low density, e.g. on aporous membrane, these substrate-bound collections are typicallydenominated macroarrays; at higher density, typically on a solidsupport, such as glass, these substrate bound collections of pluralnucleic acids are colloquially termed microarrays. As used herein, theterm microarray includes arrays of all densities. It is, therefore,another aspect of the invention to provide microarrays that include thenucleic acids of the present invention.

[0154] Expression Vectors, Host Cells and Recombinant Methods ofProducing Polypeptides

[0155] Another aspect of the present invention relates to vectors thatcomprise one or more of the isolated nucleic acid molecules of thepresent invention, and host cells in which such vectors have beenintroduced.

[0156] The vectors can be used, inter alia, for propagating the nucleicacids of the present invention in host cells (cloning vectors), forshuttling the nucleic acids of the present invention between host cellsderived from disparate organisms (shuttle vectors), for inserting thenucleic acids of the present invention into host cell chromosomes(insertion vectors), for expressing sense or antisense RNA transcriptsof the nucleic acids of the present invention in vitro or within a hostcell, and for expressing polypeptides encoded by the nucleic acids ofthe present invention, alone or as fusions to heterologous polypeptides(expression vectors). Vectors of the present invention will often besuitable for several such uses.

[0157] Vectors are by now well-known in the art, and are described,inter alia, in Jones et al. (eds.), Vectors: Cloning Applications:Essential Techniques (Essential Techniques Series), John Wiley & SonLtd. (1998); Jones et al. (eds.), Vectors: Expression Systems: EssentialTechniques (Essential Techniques Series), John Wiley & Son Ltd. (1998);Gacesa et al., Vectors: Essential Data, John Wiley & Sons Ltd. (1995);Cid-Arregui (eds.), Viral Vectors: Basic Science and Gene Therapy, EatonPublishing Co. (2000); Sambrook (2001), supra; Ausubel (1999), supra;the disclosures of which are incorporated herein by reference in theirentireties. Furthermore, an enormous variety of vectors are availablecommercially. Use of existing vectors and modifications thereof beingwell within the skill in the art, only basic features need be describedhere.

[0158] Nucleic acid sequences may be expressed by operatively linkingthem to an expression control sequence in an appropriate expressionvector and employing that expression vector to transform an appropriateunicellular host. Expression control sequences are sequences whichcontrol the transcription, post-transcriptional events and translationof nucleic acid sequences. Such operative linking of a nucleic sequenceof this invention to an expression control sequence, of course,includes, if not already part of the nucleic acid sequence, theprovision of a translation initiation codon, ATG or GTG, in the correctreading frame upstream of the nucleic acid sequence.

[0159] A wide variety of host/expression vector combinations may beemployed in expressing the nucleic acid sequences of this invention.Useful expression vectors, for example, may consist of segments ofchromosomal, non-chromosomal and synthetic nucleic acid sequences.

[0160] In one embodiment, prokaryotic cells may be used with anappropriate vector. Prokaryotic host cells are often used for cloningand expression. In a preferred embodiment, prokaryotic host cellsinclude E. coli, Pseudomonas, Bacillus and Streptomyces. In a preferredembodiment, bacterial host cells are used to express the nucleic acidmolecules of the instant invention. Useful expression vectors forbacterial hosts include bacterial plasmids, such as those from E. coli,Bacillus or Streptomyces, including pBluescript, pGEX-2T, pUC vectors,col E1, pCR1, pBR322, pMB9 and their derivatives, wider host rangeplasmids, such as RP4, phage DNAs, e.g., the numerous derivatives ofphage lambda, e.g., NM989, λGT10 and λGT11, and other phages, e.g., M13and filamentous single-stranded phage DNA. Where E. coli is used ashost, selectable markers are, analogously, chosen for selectivity ingram negative bacteria: e.g., typical markers confer resistance toantibiotics, such as ampicillin, tetracycline, chloramphenicol,kanamycin, streptomycin and zeocin; auxotrophic markers can also beused.

[0161] In other embodiments, eukaryotic host cells, such as yeast,insect, mammalian or plant cells, may be used. Yeast cells, typically S.cerevisiae, are useful for eukaryotic genetic studies, due to the easeof targeting genetic changes by homologous recombination and the abilityto easily complement genetic defects using recombinantly expressedproteins. Yeast cells are useful for identifying interacting proteincomponents, e.g. through use of a two-hybrid system. In a preferredembodiment, yeast cells are useful for protein expression. Vectors ofthe present invention for use in yeast will typically, but notinvariably, contain an origin of replication suitable for use in yeastand a selectable marker that is functional in yeast. Yeast vectorsinclude Yeast Integrating plasmids (e.g., YIp5) and Yeast Replicatingplasmids (the YRp and YEp series plasmids), Yeast Centromere plasmids(the YCp series plasmids), Yeast Artificial Chromosomes (YACs) which arebased on yeast linear plasmids, denoted YLp, pGPD-2, 2μ plasmids andderivatives thereof, and improved shuttle vectors such as thosedescribed in Gietz et al., Gene, 74: 527-34 (1988) (YIplac, YEplac andYCplac). Selectable markers in yeast vectors include a variety ofauxotrophic markers, the most common of which are (in Saccharomycescerevisiae) URA3, HIS3, LEU2, TRP1 and LYS2, which complement specificauxotrophic mutations, such as ura3-52, his3-D1, leu2-D1, trp1-D1 andlys2-201.

[0162] Insect cells are often chosen for high efficiency proteinexpression. Where the host cells are from Spodoptera frugiperda , e.g.,Sf9 and Sf21 cell lines, and expresSF™ cells (Protein Sciences Corp.,Meriden, Conn., USA)), the vector replicative strategy is typicallybased upon the baculovirus life cycle. Typically, baculovirus transfervectors are used to replace the wild-type AcMNPV polyhedrin gene with aheterologous gene of interest. Sequences that flank the polyhedrin genein the wild-type genome are positioned 5′ and 3′ of the expressioncassette on the transfer vectors. Following co-transfection with AcMNPVDNA, a homologous recombination event occurs between these sequencesresulting in a recombinant virus carrying the gene of interest and thepolyhedrin or p10 promoter. Selection can be based upon visual screeningfor lacZ fusion activity.

[0163] In another embodiment, the host cells may be mammalian cells,which are particularly useful for expression of proteins intended aspharmaceutical agents, and for screening of potential agonists andantagonists of a protein or a physiological pathway. Mammalian vectorsintended for autonomous extrachromosomal replication will typicallyinclude a viral origin, such as the SV40 origin (for replication in celllines expressing the large T-antigen, such as COSI and COS7 cells), thepapillomavirus origin, or the EBV origin for long term episomalreplication (for use, e.g., in 293-EBNA cells, which constitutivelyexpress the EBV EBNA-1 gene product and adenovirus E1A). Vectorsintended for integration, and thus replication as part of the mammalianchromosome, can, but need not, include an origin of replicationfunctional in mammalian cells, such as the SV40 origin. Vectors basedupon viruses, such as adenovirus, adeno-associated virus, vacciniavirus, and various mammalian retroviruses, will typically replicateaccording to the viral replicative strategy. Selectable markers for usein mammalian cells include resistance to neomycin (G418), blasticidin,hygromycin and to zeocin, and selection based upon the purine salvagepathway using HAT medium.

[0164] Expression in mammalian cells can be achieved using a variety ofplasmids, including pSV2, pBC12BI, and p91023, as well as lytic virusvectors (e.g., vaccinia virus, adeno virus, and baculovirus), episomalvirus vectors (e.g., bovine papillomavirus), and retroviral vectors(e.g., murine retroviruses). Useful vectors for insect cells includebaculoviral vectors and pVL 941.

[0165] Plant cells can also be used for expression, with the vectorreplicon typically derived from a plant virus (e.g., cauliflower mosaicvirus, CaMV; tobacco mosaic virus, TMV) and selectable markers chosenfor suitability in plants.

[0166] It is known that codon usage of different host cells may bedifferent. For example, a plant cell and a human cell may exhibit adifference in codon preference for encoding a particular amino acid. Asa result, human mRNA may not be efficiently translated in a plant,bacteria or insect host cell. Therefore, another embodiment of thisinvention is directed to codon optimization. The codons of the nucleicacid molecules of the invention may be modified to resemble, as much aspossible, genes naturally contained within the host cell withoutaltering the amino acid sequence encoded by the nucleic acid molecule.

[0167] Any of a wide variety of expression control sequences may be usedin these vectors to express the DNA sequences of this invention. Suchuseful expression control sequences include the expression controlsequences associated with structural genes of the foregoing expressionvectors. Expression control sequences that control transcriptioninclude, e.g., promoters, enhancers and transcription termination sites.Expression control sequences in eukaryotic cells that controlpost-transcriptional events include splice donor and acceptor sites andsequences that modify the half-life of the transcribed RNA, e.g.,sequences that direct poly(A) addition or binding sites for RNA-bindingproteins. Expression control sequences that control translation includeribosome binding sites, sequences which direct targeted cxprcssion ofthe polypeptide to or within particular cellular compartments, andsequences in the 5′ and 3′ untranslated regions that modify the rate orefficiency of translation.

[0168] Examples of useful expression control sequences for a prokaryote,e.g., E. coli, will include a promoter, often a phage promoter, such asphage lambda pL promoter, the trc promoter, a hybrid derived from thetrp and lac promoters, the bacteriophage T7 promoter (in E. coli cellsengineered to express the T7 polymerase), the TAC or TRC system, themajor operator and promoter regions of phage lambda, the control regionsof fd coat protein, or the araBAD operon. Prokaryotic expression vectorsmay further include transcription terminators, such as the aspAterminator, and elements that facilitate translation, such as aconsensus ribosome binding site and translation termination codon,Schomer et al., Proc. Natl. Acad. Sci. USA 83: 8506-8510 (1986).

[0169] Expression control sequences for yeast cells, typically S.cerevisiae, will include a yeast promoter, such as the CYC1 promoter,the GAL1 promoter, the GAL10 promoter, ADH1 promoter, the promoters ofthe yeast_-mating system, or the GPD promoter, and will typically haveelements that facilitate transcription termination, such as thetranscription termination signals from the CYC1 or ADH1 gene.

[0170] Expression vectors useful for expressing proteins in mammaliancells will include a promoter active in mammalian cells. These promotersinclude those derived from mammalian viruses, such as theenhancer-promoter sequences from the immediate early gene of the humancytomegalovirus (CMV), the enhancer-promoter sequences from the Roussarcoma virus long terminal repeat (RSV LTR), the enhancer-promoter fromSV40 or the early and late promoters of adenovirus. Other expressioncontrol sequences include the promoter for 3-phosphoglycerate kinase orother glycolytic enzymes, the promoters of acid phosphatase. Otherexpression control sequences include those from the gene comprising theBSNA of interest. Often, expression is enhanced by incorporation ofpolyadenylation sites, such as the late SV40 polyadenylation site andthe polyadenylation signal and transcription termination sequences fromthe bovine growth hormone (BGH) gene, and ribosome binding sites.Furthermore, vectors can include introns, such as intron II of rabbitβ-globin gene and the SV40 splice elements.

[0171] Preferred nucleic acid vectors also include a selectable oramplifiable marker gene and means for amplifying the copy number of thegene of interest. Such marker genes are well-known in the art. Nucleicacid vectors may also comprise stabilizing sequences (e.g., ori- orARS-like sequences and telomere-like sequences), or may alternatively bedesigned to favor directed or non-directed integration into the hostcell genome. In a preferred embodiment, nucleic acid sequences of thisinvention are inserted in frame into an expression vector that allowshigh level expression of an RNA which encodes a protein comprising theencoded nucleic acid sequence of interest. Nucleic acid cloning andsequencing methods are well-known to those of skill in the art and aredescribed in an assortment of laboratory manuals, including Sambrook(1989), supra, Sambrook (2000), supra; and Ausubel (1 992), supra,Ausubel (1999), supra. Product information from manufacturers ofbiological, chemical and immunological reagents also provide usefulinformation.

[0172] Expression vectors may be either constitutive or inducible.Inducible vectors include either naturally inducible promoters, such asthe trc promoter, which is regulated by the lac operon, and the pLpromoter, which is regulated by tryptophan, the MMTV-LTR promoter, whichis inducible by dexamethasone, or can contain synthetic promoters and/oradditional elements that confer inducible control on adjacent promoters.Examples of inducible synthetic promoters are the hybrid Plac/ara-1promoter and the PLtetO-1 promoter. The PltetO-1 promoter takesadvantage of the high expression levels from the PL promoter of phagelambda, but replaces the lambda repressor sites with two copies ofoperator 2 of the Tn10 tetracycline resistance operon, causing thispromoter to be tightly repressed by the Tet repressor protein andinduced in response to tetracycline (Tc) and Tc derivatives such asanhydrotetracycline. Vectors may also be inducible because they containhormone response elements, such as the glucocorticoid response element(GRE) and the estrogen response element (ERE), which can confer hormoneinducibility where vectors are used for expression in cells having therespective hormone receptors. To reduce background levels of expression,elements responsive to ecdysone, an insect hormone, can be used instead,with coexpression of the ecdysone receptor.

[0173] In one aspect of the invention, expression vectors can bedesigned to fuse the expressed polypeptide to small protein tags thatfacilitate purification and/or visualization. Tags that facilitatepurification include a polyhistidine tag that facilitates purificationof the fusion protein by immobilized metal affinity chromatography, forexample using NiNTA resin (Qiagen Inc., Valencia, Calif., USA) or TALON™resin (cobalt immobilized affinity chromatography medium, Clontech Labs,Palo Alto, Calif., USA). The fusion protein can include a chitin-bindingtag and self-excising intein, permitting chitin-based purification withself-removal of the fused tag (IMPACT™ system, New England Biolabs,Inc., Beverley, Mass., USA). Alternatively, the fusion protein caninclude a calmodulin-binding peptide tag, permitting purification bycalmodulin affinity resin (Stratagene, La Jolla, Calif., USA), or aspecifically excisable fragment of the biotin carboxylase carrierprotein, permitting purification of in vivo biotinylated protein usingan avidin resin and subsequent tag removal (Promega, Madison, Wis.,USA). As another useful alternative, the proteins of the presentinvention can be expressed as a fusion protein withglutathione-S-transferase, the affinity and specificity of binding toglutathione permitting purification using glutathione affinity resins,such as Glutathione-Superflow Resin (Clontech Laboratories, Palo Alto,Calif., USA), with subsequent elution with free glutathione. Other tagsinclude, for example, the Xpress epitope, detectable by anti-Xpressantibody (Invitrogen, Carlsbad, Calif., USA), a myc tag, detectable byanti-myc tag antibody, the V5 epitope, detectable by anti-V5 antibody(Invitrogen, Carlsbad, Calif., USA), FLAG® epitope, detectable byanti-FLAG® antibody (Stratagene, La Jolla, Calif., USA), and the HAepitope.

[0174] For secretion of expressed proteins, vectors can includeappropriate sequences that encode secretion signals, such as leaderpeptides. For example, the pSecTag2 vectors (Invitrogen, Carlsbad,Calif., USA) are 5.2 kb mammalian expression vectors that carry thesecretion signal from the V-J2-C region of the mouse Ig kappa-chain forefficient secretion of recombinant proteins from a variety of mammaliancell lines.

[0175] Expression vectors can also be designed to fuse proteins encodedby the heterologous nucleic acid insert to polypeptides that are largerthan purification and/or identification tags. Useful fusion proteinsinclude those that permit display of the encoded protein on the surfaceof a phage or cell, fusion to intrinsically fluorescent proteins, suchas those that have a green fluorescent protein (GFP)-like chromophore,fusions to the IgG Fc region, and fusion proteins for use in two hybridsystems.

[0176] Vectors for phage display fuse the encoded polypeptide to, e.g.,the gene III protein (pIII) or gene VIII protein (pVIII) for display onthe surface of filamentous phage, such as M13. See Barbas et al., PhageDisplay: A Laboratory Manual, Cold Spring Harbor Laboratory Press(2001); Kay et al. (eds.), Phage Display of Peptides and Proteins: ALaboratory Manual, Academic Press, Inc., (1996); Abelson et al. (eds.),Combinatorial Chemistry (Methods in Enzymology, Vol. 267) Academic Press(1996). Vectors for yeast display, e.g. the pYD1 yeast display vector(Invitrogen, Carlsbad, Calif., USA), use the -agglutinin yeast adhesionreceptor to display recombinant protein on the surface of S. cerevisiae.Vectors for mammalian display, e.g., the pDisplay™ vector (Invitrogen,Carlsbad, Calif., USA), target recombinant proteins using an N-terminalcell surface targeting signal and a C-terminal transmembrane anchoringdomain of platelet derived growth factor receptor.

[0177] A wide variety of vectors now exist that fuse proteins encoded byheterologous nucleic acids to the chromophore of thesubstrate-independent, intrinsically fluorescent green fluorescentprotein from Aequorea victoria (“GFP”) and its variants. The GFP-likechromophore can be selected from GFP-like chromophores found innaturally occurring proteins, such as A. victoria GFP (GenBank accessionnumber AAA27721), Renilla reniformis GFP, FP583 (GenBank accession no.AF168419) (DsRed), FP593 (AF27271 1), FP483 (AF168420), FP484(AF168424), FP595 (AF246709), FP486 (AF168421), FP538 (AF168423), andFP506 (AF168422), and need include only so much of the native protein asis needed to retain the chromophore's intrinsic fluorescence. Methodsfor determining the minimal domain required for fluorescence are knownin the art. See Li et al., J. Biol. Chem. 272: 28545-28549 (1997).Alternatively, the GFP-like chromophore can be selected from GFP-likechromophores modified from those found in nature. The methods forengineering such modified GFP-like chromophores and testing them forfluorescence activity, both alone and as part of protein fusions, arewell-known in the art. See Heim et al., Curr. Biol. 6: 178-182 (1996)and Palm et al., Methods Enzymol. 302: 378-394 (1999), incorporatedherein by reference in its entirety. A variety of such modifiedchromophores are now commercially available and can readily be used inthe fusion proteins of the present invention. These include EGFP(“enhanced GFP”), EBFP (“enhanced blue fluorescent protein”), BFP2, EYFP(“enhanced yellow fluorescent protein”), ECFP (“enhanced cyanfluorescent protein”) or Citrine. EGFP (see, e.g. Cormack et al., Gene173: 33-38 (1996); U.S. Pat. Nos. 6,090,919 and 5,804,387) is found on avariety of vectors, both plasmid and viral, which are availablecommercially (Clontech Labs, Palo Alto, Calif., USA); EBFP is optimizedfor expression in mammalian cells whereas BFP2, which retains theoriginal jellyfish codons, can be expressed in bacteria (see, e.g,. Heimet al., Curr. Biol. 6: 178-182 (1996) and Cormack et al., Gene 173:33-38 (1996)). Vectors containing these blue-shifted variants areavailable from Clontech Labs (Palo Alto, Calif., USA). Vectorscontaining EYFP, ECFP (see, e.g., Heim et al., Curr. Biol. 6: 178-182(1996); Miyawaki et al., Nature 388: 882-887 (1997)) and Citrine (see,e.g., Heikal et al., Proc. Natl. Acad. Sci. USA 97: 11996-12001 (2000))are also available from Clontech Labs. The GFP-like chromophore can alsobe drawn from other modified GFPs, including those described in U.S.Pat. Nos. 6,124,128; 6,096,865; 6,090,919; 6,066,476; 6,054,321;6,027,881; 5,968,750; 5,874,304; 5,804,387; 5,777,079; 5,741,668; and5,625,048, the disclosures of which are incorporated herein by referencein their entireties. See also Conn (ed.), Green Fluorescent Protein(Methods in Enzymology, Vol. 302), Academic Press, Inc. (1999). TheGFP-like chromophore of each of these GFP variants can usefully beincluded in the fusion proteins of the present invention.

[0178] Fusions to the IgG Fc region increase serum half life of proteinpharmaceutical products through interaction with the FcRn receptor (alsodenominated the FcRp receptor and the Brambell receptor, FcRb), furtherdescribed in International Patent Application Nos. WO 97/43316, WO97/34631, WO 96/32478, WO 96/18412.

[0179] For long-term, high-yield recombinant production of the proteins,protein fusions, and protein fragments of the present invention, stableexpression is preferred. Stable expression is readily achieved byintegration into the host cell genome of vectors having selectablemarkers, followed by selection of these integrants. Vectors such aspUB6/V5-His A, B, and C (Invitrogen, Carlsbad, Calif., USA) are designedfor high-level stable expression of heterologous proteins in a widerange of mammalian tissue types and cell lines. pUB6/V5-His uses thepromoter/enhancer sequence from the human ubiquitin C gene to driveexpression of recombinant proteins: expression levels in 293, CHO, andNIH3T3 cells are comparable to levels from the CMV and human EF-1apromoters. The bsd gene permits rapid selection of stably transfectedmammalian cells with the potent antibiotic blasticidin.

[0180] Replication incompetent retroviral vectors, typically derivedfrom Moloney murine leukemia virus, also are useful for creating stabletransfectants having integrated provirus. The highly efficienttransduction machinery of retroviruses, coupled with the availability ofa variety of packaging cell lines such as RetroPack™ PT 67, EcoPack2™-293, AmphoPack-293, and GP2-293 cell lines (all available from ClontechLaboratories, Palo Alto, Calif., USA), allow a wide host range to beinfected with high efficiency; varying the multiplicity of infectionreadily adjusts the copy number of the integrated provirus.

[0181] Of course, not all vectors and expression control sequences willfunction equally well to express the nucleic acid sequences of thisinvention. Neither will all hosts function equally well with the sameexpression system. However, one of skill in the art may make a selectionamong these vectors, expression control sequences and hosts withoutundue experimentation and without departing from the scope of thisinvention. For example, in selecting a vector, the host must beconsidered because the vector must be replicated in it. The vector'scopy number, the ability to control that copy number, the ability tocontrol integration, if any, and the expression of any other proteinsencoded by the vector, such as antibiotic or other selection markers,should also be considered. The present invention further includes hostcells comprising the vectors of the present invention, either presentepisomally within the cell or integrated, in whole or in part, into thehost cell chromosome. Among other considerations, some of which aredescribed above, a host cell strain may be chosen for its ability toprocess the expressed protein in the desired fashion. Suchpost-translational modifications of the polypeptide include, but are notlimited to, acetylation, carboxylation, glycosylation, phosphorylation,lipidation, and acylation, and it is an aspect of the present inventionto provide BSPs with such post-translational modifications.

[0182] Polypeptides of the invention may be post-translationallymodified. Post-translational modifications include phosphorylation ofamino acid residues serine, threonine and/or tyrosine, N-linked and/orO-linked glycosylation, methylation, acetylation, prenylation,methylation, acetylation, arginylation, ubiquination and racemization.One may determine whether a polypeptide of the invention is likely to bepost-translationally modified by analyzing the sequence of thepolypeptide to determine if there are peptide motifs indicative of sitesfor post-translational modification. There are a number of computerprograms that permit prediction of post-translational modifications.See, e.g., www.expasy.org (accessed Aug. 31, 2001), which includesPSORT, for prediction of protein sorting signals and localization sites,SignalP, for prediction of signal peptide cleavage sites, MITOPROT andPredotar, for prediction of mitochondrial targeting sequences, NetOGlyc,for prediction of type O-glycosylation sites in mammalian proteins,big-PI Predictor and DGPI, for prediction of prenylation-anchor andcleavage sites, and NetPhos, for prediction of Ser, Thr and Tyrphosphorylation sites in eukaryotic proteins. Other computer programs,such as those included in GCG, also may be used to determinepost-translational modification peptide motifs.

[0183] General examples of types of post-translational modifications maybe found in web sites such as the Delta Mass databasehttp://www.abrf.org/ABRF/Research Committees/deltamass/deltamass.html(accessed Oct. 19, 2001); “GlycoSuiteDB: a new curated relationaldatabase of glycoprotein glycan structures and their biological sources”Cooper et al. Nucleic Acids Res. 29; 332-335 (2001) andhttp://www.glycosuite.com/ (accessed Oct. 19, 2001); “O-GLYCBASE version4.0: a revised database of O-glycosylated proteins” Gupta et al. NucleicAcids Research, 27: 370-372 (1999) andhttp://www.cbs.dtu.dk/databases/OCLYCBASE/ (accessed Oct. 19, 2001);“PhosphoBase, a database of phosphorylation sites: release 2.0.”,Kreegipuu et al. Nucleic Acids Res 27(1):237-239 (1999) andhttp://www.cbs.dtu.dk/databases/PhosphoBase/ (accessed Oct. 19, 2001);or http://pir.georgetown.edu/pirwww/search/textresid.html (accessed Oct.19, 2001).

[0184] Tumorigenesis is often accompanied by alterations in thepost-translational modifications of proteins. Thus, in anotherembodiment, the invention provides polypeptides from cancerous cells ortissues that have altered post-translational modifications compared tothe post-translational modifications of polypeptides from normal cellsor tissues. A number of altered post-translational modifications areknown. One common alteration is a change in phosphorylation state,wherein the polypeptide from the cancerous cell or tissue ishyperphosphorylated or hypophosphorylated compared to the polypeptidefrom a normal tissue, or wherein the polypeptide is phosphorylated ondifferent residues than the polypeptide from a normal cell. Anothercommon alteration is a change in glycosylation state, wherein thepolypeptide from the cancerous cell or tissue has more or lessglycosylation than the polypeptide from a normal tissue, and/or whereinthe polypeptide from the cancerous cell or tissue has a different typeof glycosylation than the polypeptide from a noncancerous cell ortissue. Changes in glycosylation may be critical becausecarbohydrate-protein and carbohydrate-carbohydrate interactions areimportant in cancer cell progression, dissemination and invasion. See,e.g., Barchi, Curr. Pharm. Des. 6: 485-501 (2000), Verma, CancerBiochem. Biophys. 14: 151-162 (1994) and Dennis et al., Bioessays 5:412-421 (1999).

[0185] Another post-translational modification that may be altered incancer cells is prenylation. Prenylation is the covalent attachment of ahydrophobic prenyl group (either farnesyl or geranylgeranyl) to apolypeptide. Prenylation is required for localizing a protein to a cellmembrane and is often required for polypeptide function. For instance,the Ras superfamily of GTPase signaling proteins must be prenylated forfunction in a cell. See, e.g., Prendergast et al., Semin. Cancer Biol.10: 443-452 (2000) and Khwaja et al., Lancet 355: 741-744 (2000).

[0186] Other post-translation modifications that may be altered incancer cells include, without limitation, polypeptide methylation,acetylation, arginylation or racemization of amino acid residues. Inthese cases, the polypeptide from the cancerous cell may exhibit eitherincreased or decreased amounts of the post-translational modificationcompared to the corresponding polypeptides from noncancerous cells.

[0187] Other polypeptide alterations in cancer cells include abnormalpolypeptide cleavage of proteins and aberrant protein-proteininteractions. Abnormal polypeptide cleavage may be cleavage of apolypeptide in a cancerous cell that does not usually occur in a normalcell, or a lack of cleavage in a cancerous cell, wherein the polypeptideis cleaved in a normal cell. Aberrant protein-protein interactions maybe either covalent cross-linking or non-covalent binding betweenproteins that do not normally bind to each other. Alternatively, in acancerous cell, a protein may fail to bind to another protein to whichit is bound in a noncancerous cell. Alterations in cleavage or inprotein-protein interactions may be due to over- or underproduction of apolypeptide in a cancerous cell compared to that in a normal cell, ormay be due to alterations in post-translational modifications (seeabove) of one or more proteins in the cancerous cell. See, e.g.,Henschen-Edman, Ann. N.Y. Acad. Sci. 936: 580-593 (2001).

[0188] Alterations in polypeptide post-translational modifications, aswell as changes in polypeptide cleavage and protein-proteininteractions, may be determined by any method known in the art. Forinstance, alterations in phosphorylation may be determined by usinganti-phosphoserine, anti-phosphothreonine or anti-phosphotyrosineantibodies or by amino acid analysis. Glycosylation alterations may bedetermined using antibodies specific for different sugar residues, bycarbohydrate sequencing, or by alterations in the size of theglycoprotein, which can be determined by, e.g., SDS polyacrylamide gelelectrophoresis (PAGE). Other alterations of post-translationalmodifications, such as prenylation, racemization, methylation,acetylation and arginylation, may be determined by chemical analysis,protein sequencing, amino acid analysis, or by using antibodies specificfor the particular post-translational modifications. Changes inprotein-protein interactions and in polypeptide cleavage may be analyzedby any method known in the art including, without limitation,non-denaturing PAGE (for non-covalent protein-protein interactions), SDSPAGE (for covalent protein-protein interactions and protein cleavage),chemical cleavage, protein sequencing or immunoassays.

[0189] In another embodiment, the invention provides polypeptides thathave been post-translationally modified. In one embodiment, polypeptidesmay be modified enzymatically or chemically, by addition or removal of apost-translational modification. For example, a polypeptide may beglycosylated or deglycosylated enzymatically. Similarly, polypeptidesmay be phosphorylated using a purified kinase, such as a MAP kinase(e.g, p38, ERK, or JNK) or a tyrosine kinase (e.g., Src or erbB2). Apolypeptide may also be modified through synthetic chemistry.Alternatively, one may isolate the polypeptide of interest from a cellor tissue that expresses the polypeptide with the desiredpost-translational modification. In another embodiment, a nucleic acidmolecule encoding the polypeptide of interest is introduced into a hostcell that is capable of post-translationally modifying the encodedpolypeptide in the desired fashion. If the polypeptide does not containa motif for a desired post-translational modification, one may alter thepost-translational modification by mutating the nucleic acid sequence ofa nucleic acid molecule encoding the polypeptide so that it contains asite for the desired post-translational modification. Amino acidsequences that may be post-translationally modified are known in theart. See, e.g., the programs described above on the websitewww.expasy.org. The nucleic acid molecule is then be introduced into ahost cell that is capable of post-translationally modifying the encodedpolypeptide. Similarly, one may delete sites that arepost-translationally modified by either mutating the nucleic acidsequence so that the encoded polypeptide does not contain thepost-translational modification motif, or by introducing the nativenucleic acid molecule into a host cell that is not capable ofpost-translationally modifying the encoded polypeptide.

[0190] In selecting an expression control sequence, a variety of factorsshould also be considered. These include, for example, the relativestrength of the sequence, its controllability, and its compatibilitywith the nucleic acid sequence of this invention, particularly withregard to potential secondary structures. Unicellular hosts should beselected by consideration of their compatibility with the chosen vector,the toxicity of the product coded for by the nucleic acid sequences ofthis invention, their secretion characteristics, their ability to foldthe polypeptide correctly, their fermentation or culture requirements,and the ease of purification from them of the products coded for by thenucleic acid sequences of this invention.

[0191] The recombinant nucleic acid molecules and more particularly, theexpression vectors of this invention may be used to express thepolypeptides of this invention as recombinant polypeptides in aheterologous host cell. The polypeptides of this invention may befull-length or less than full-length polypeptide fragments rccombinantlyexpressed from the nucleic acid sequences according to this invention.Such polypeptides include analogs, derivatives and muteins that may ormay not have biological activity.

[0192] Vectors of the present invention will also often include elementsthat permit in vitro transcription of RNA from the inserted heterologousnucleic acid. Such vectors typically include a phage promoter, such asthat from T7, T3, or SP6, flanking the nucleic acid insert. Often twodifferent such promoters flank the inserted nucleic acid, permittingseparate in vitro production of both sense and antisense strands.

[0193] Transformation and other methods of introducing nucleic acidsinto a host cell (e.g., conjugation, protoplast transformation orfusion, transfection, electroporation, liposome delivery, membranefusion techniques, high velocity DNA-coated pellets, viral infection andprotoplast fusion) can be accomplished by a variety of methods which arewell-known in the art (See, for instance, Ausubel, supra, and Sambrooket al., supra). Bacterial, yeast, plant or mammalian cells aretransformed or transfected with an expression vector, such as a plasmid,a cosmid, or the like, wherein the expression vector comprises thenucleic acid of interest. Alternatively, the cells may be infected by aviral expression vector comprising the nucleic acid of interest.Depending upon the host cell, vector, and method of transformation used,transient or stable expression of the polypeptide will be constitutiveor inducible. One having ordinary skill in the art will be able todecide whether to express a polypeptide transiently or stably, andwhether to express the protein constitutively or inducibly.

[0194] A wide variety of unicellular host cells are useful in expressingthe DNA sequences of this invention. These hosts may include well-knowneukaryotic and prokaryotic hosts, such as strains of, fungi, yeast,insect cells such as Spodoptera frugiperda (SF9), animal cells such asCHO, as well as plant cells in tissue culture. Representative examplesof appropriate host cells include, but are not limited to, bacterialcells, such as E. coli, Caulobacter crescentus, Streptomyces species,and Salmonella typhimurium; yeast cells, such as Saccharomycescerevisiae, Schizosaccharomyces pombe, Pichia pastoris, Pichiamethanolica; insect cell lines, such as those from Spodopterafrugiperda, e.g., Sf9 and Sf21 cell lines, and expresSF™ cells (ProteinSciences Corp., Meriden, Conn., USA), Drosophila S2 cells, andTrichoplusia ni High Five® Cells (Invitrogen, Carlsbad, Calif., USA);and mammalian cells. Typical mammalian cells include BHK cells, BSC 1cells, BSC 40 cells, BMT 10 cells, VERO cells, COS1 cells, COS7 cells,Chinese hamster ovary (CHO) cells, 3T3 cells, NIH 3T3 cells, 293 cells,HEPG2 cells, HeLa cells, L cells, MDCK cells, HEK293 cells, W138 cells,murine ES cell lines (e.g., from strains 129/SV, C57/BL6, DBA-1,129/SVJ), K562 cells, Jurkat cells, and BW5147 cells. Other mammaliancell lines are well-known and readily available from the American TypeCulture Collection (ATCC) (Manassas, Va., USA) and the NationalInstitute of General Medical Sciences (NIGMS) Human Genetic CellRepository at the Coriell Cell Repositories (Camden, N.J., USA). Cellsor cell lines derived from breast are particularly preferred becausethey may provide a more native post-translational processing.Particularly preferred are human breast cells.

[0195] Particular details of the transfection, expression andpurification of recombinant proteins are well documented and areunderstood by those of skill in the art. Further details on the varioustechnical aspects of each of the steps used in recombinant production offoreign genes in bacterial cell expression systems can be found in anumber of texts and laboratory manuals in the art. See, e.g., Ausubel(1992), supra, Ausubel (1999), supra, Sambrook (1989), supra, andSambrook (2001), supra, herein incorporated by reference.

[0196] Methods for introducing the vectors and nucleic acids of thepresent invention into the host cells are well-known in the art; thechoice of technique will depend primarily upon the specific vector to beintroduced and the host cell chosen.

[0197] Nucleic acid molecules and vectors may be introduced intoprokaryotes, such as E. coli, in a number of ways. For instance, phagelambda vectors will typically be packaged using a packaging extract(e.g., Gigapack® packaging extract, Stratagene, La Jolla, Calif., USA),and the packaged virus used to infect E. coli.

[0198] Plasmid vectors will typically be introduced into chemicallycompetent or electrocompetent bacterial cells. E. coli cells can berendered chemically competent by treatment, e.g., with CaCl₂, or asolution of Mg²⁺, Mn²⁺, Ca²⁺, Rb⁺ or K⁺, dimethyl sulfoxide,dithiothreitol, and hexamine cobalt (III), Hanahan, J. Mol. Biol.166(4):557-80 (1983), and vectors introduced by heat shock. A widevariety of chemically competent strains are also available commercially(e.g., Epicurian Coli® XL10-Gold® Ultracompetent Cells (Stratagene, LaJolla, Calif., USA); DH5 competent cells (Clontech Laboratories, PaloAlto, Calif., USA); and TOP10 Chemically Competent E. coli Kit(Invitrogen, Carlsbad, Calif., USA)). Bacterial cells can be renderedelectrocompetent, that is, competent to take up exogenous DNA byelectroporation, by various pre-pulse treatments; vectors are introducedby electroporation followed by subsequent outgrowth in selected media.An extensive series of protocols is provided online in Electroprotocols(BioRad, Richmond, Calif., USA)(http://www.biorad.com/LifeScience/pdf/New_Gene_Pulser.pdf).

[0199] Vectors can be introduced into yeast cells by spheroplasting,treatment with lithium salts, electroporation, or protoplast fusion.Spheroplasts are prepared by the action of hydrolytic enzymes such assnail-gut extract, usually denoted Glusulase, or Zymolyase, an enzymefrom Arthrobacter luteus, to remove portions of the cell wall in thepresence of osmotic stabilizers, typically 1 M sorbitol. DNA is added tothe spheroplasts, and the mixture is co-precipitated with a solution ofpolyethylene glycol (PEG) and Ca²⁺. Subsequently, the cells areresuspended in a solution of sorbitol, mixed with molten agar and thenlayered on the surface of a selective plate containing sorbitol.

[0200] For lithium-mediated transformation, yeast cells are treated withlithium acetate, which apparently permeabilizes the cell wall, DNA isadded and the cells are co-precipitated with PEG. The cells are exposedto a brief heat shock, washed free of PEG and lithium acetate, andsubsequently spread on plates containing ordinary selective medium.Increased frequencies of transformation are obtained by usingspecially-prepared single-stranded carrier DNA and certain organicsolvents. Schiestl et al., Curr. Genet. 16(5-6): 339-46 (1989).

[0201] For electroporation, freshly-grown yeast cultures are typicallywashed, suspended in an osmotic protectant, such as sorbitol, mixed withDNA, and the cell suspension pulsed in an electroporation device.Subsequently, the cells are spread on the surface of plates containingselective media. Becker et al., Methods Enzymol. 194: 182-187 (1991).The efficiency of transformation by electroporation can be increasedover 100-fold by using PEG, single-stranded carrier DNA and cells thatare in late log-phase of growth. Larger constructs, such as YACs, can beintroduced by protoplast fusion.

[0202] Mammalian and insect cells can be directly infected by packagedviral vectors, or transfected by chemical or electrical means. Forchemical transfection, DNA can be coprecipitated with CaPO₄ orintroduced using liposomal and nonliposomal lipid-based agents.Commercial kits are available for CaPO₄ transfection (CalPhos™ MammalianTransfection Kit, Clontech Laboratories, Palo Alto, Calif., USA), andlipid-mediated transfection can be practiced using commercial reagents,such as LIPOFECTAMINE™ 2000, LIPOFECTAMINE™ Reagent, CELLFECTIN™Reagent, and LIPOFECTIN™ Reagent (Invitrogen, Carlsbad, Calif., USA),DOTAP Liposomal Transfection Reagent, FuGENE 6, X-tremeGENE Q2, DOSPER,(Roche Molecular Biochemicals, Indianapolis, Ind. USA), Effectene™,PolyFect®, Superfect® (Qiagen, Inc., Valencia, Calif., USA). Protocolsfor electroporating mammalian cells can be found online inElectroprotocols (Bio-Rad, Richmond, Calif., USA)(http://www.bio-rad.com/LifeScience/pdf/New_Gene_Pulser.pdf); Norton etal. (eds.), Gene Transfer Methods: Introducing DNA into Living Cells andOrganisms, BioTechniques Books, Eaton Publishing Co. (2000);incorporated herein by reference in its entirety. Other transfectiontechniques include transfection by particle bombardment andmicroinjection. See, e.g., Cheng et al., Proc. Natl. Acad. Sci. USA90(10): 4455-9 (1993); Yang et al., Proc. Natl. Acad. Sci. USA 87(24):9568-72 (1990).

[0203] Production of the recombinantly produced proteins of the presentinvention can optionally be followed by purification.

[0204] Purification of recombinantly expressed proteins is now well bythose skilled in the art. See, e.g., Thorner et al. (eds.), Applicationsof Chimeric Genes and Hybrid Proteins, Part A: Gene Expression andProtein Purification (Methods in Enzymology, Vol. 326), Academic Press(2000); Harbin (ed.), Cloning, Gene Expression and Protein Purification:Experimental Procedures and Process Rationale, Oxford Univ. Press(2001); Marshak et al., Strategies for Protein Purification andCharacterization: A Laboratory Course Manual, Cold Spring HarborLaboratory Press (1996); and Roe (ed.), Protein PurificationApplications, Oxford University Press (2001); the disclosures of whichare incorporated herein by reference in their entireties, and thus neednot be detailed here.

[0205] Briefly, however, if purification tags have been fused throughuse of an expression vector that appends such tags, purification can beeffected, at least in part, by means appropriate to the tag, such as useof immobilized metal affinity chromatography for polyhistidine tags.Other techniques common in the art include ammonium sulfatefractionation, immunoprecipitation, fast protein liquid chromatography(FPLC), high performance liquid chromatography (HPLC), and preparativegel electrophoresis.

[0206] Polypeptides

[0207] Another object of the invention is to provide polypeptidesencoded by the nucleic acid molecules of the instant invention. In apreferred embodiment, the polypeptide is a breast specific polypeptide(BSP). In an even more preferred embodiment, the polypeptide is derivedfrom a polypeptide comprising the amino acid sequence of SEQ ID NO: 160through 282. A polypeptide as defined herein may be producedrecombinantly, as discussed supra, may be isolated from a cell thatnaturally expresses the protein, or may be chemically synthesizedfollowing the teachings of the specification and using methodswell-known to those having ordinary skill in the art.

[0208] In another aspect, the polypeptide may comprise a fragment of apolypeptide, wherein the fragment is as defined herein. In a preferredembodiment, the polypeptide fragment is a fragment of a BSP. In a morepreferred embodiment, the fragment is derived from a polypeptidecomprising the amino acid sequence of SEQ ID NO: 160 through 282. Apolypeptide that comprises only a fragment of an entire BSP may or maynot be a polypeptide that is also a BSP. For instance, a full-lengthpolypeptide may be breast-specific, while a fragment thereof may befound in other tissues as well as in breast. A polypeptide that is not aBSP, whether it is a fragment, analog, mutein, homologous protein orderivative, is nevertheless useful, especially for immunizing animals toprepare anti-BSP antibodies. However, in a preferred embodiment, thepart or fragment is a BSP. Methods of determining whether a polypeptideis a BSP are described infra.

[0209] Fragments of at least 6 contiguous amino acids are useful inmapping B cell and T cell epitopes of the reference protein. See, e.g.,Geysen et al., Proc. Natl. Acad. Sci. USA 81: 3998-4002 (1984) and U.S.Pat. Nos. 4,708,871 and 5,595,915, the disclosures of which areincorporated herein by reference in their entireties. Because thefragment need not itself be immunogenic, part of an immunodominantepitope, nor even recognized by native antibody, to be useful in suchepitope mapping, all fragments of at least 6 amino acids of the proteinsof the present invention have utility in such a study.

[0210] Fragments of at least 8 contiguous amino acids, often at least 15contiguous amino acids, are useful as immunogens for raising antibodiesthat recognize the proteins of the present invention. See, e.g., Lerner,Nature 299: 592-596 (1982); Shinnick et al., Annu. Rev. Microbiol. 37:425-46 (1983); Sutcliffe et al., Science 219: 660-6 (1983), thedisclosures of which are incorporated herein by reference in theirentireties. As further described in the above-cited references,virtually all 8-mers, conjugated to a carrier, such as a protein, proveimmunogenic, meaning that they are capable of eliciting antibody for theconjugated peptide; accordingly, all fragments of at least 8 amino acidsof the proteins of the present invention have utility as immunogens.

[0211] Fragments of at least 8, 9, 10 or 12 contiguous amino acids arealso useful as competitive inhibitors of binding of the entire protein,or a portion thereof, to antibodies (as in epitope mapping), and tonatural binding partners, such as subunits in a multimeric complex or toreceptors or ligands of the subject protein; this competitive inhibitionpermits identification and separation of molecules that bindspecifically to the protein of interest, U.S. Pat. Nos. 5,539,084 and5,783,674, incorporated herein by reference in their entireties.

[0212] The protein, or protein fragment, of the present invention isthus at least 6 amino acids in length, typically at least 8, 9, 10 or 12amino acids in length, and often at least 15 amino acids in length.Often, the protein of the present invention, or fragment thereof, is atleast 20 amino acids in length, even 25 amino acids, 30 amino acids, 35amino acids, or 50 amino acids or more in length. Of course, largerfragments having at least 75 amino acids, 100 amino acids, or even 150amino acids are also useful, and at times preferred.

[0213] One having ordinary skill in the art can produce fragments of apolypeptide by truncating the nucleic acid molecule, e.g., a BSNA,encoding the polypeptide and then expressing it recombinantly.Alternatively, one can produce a fragment by chemically synthesizing aportion of the full-length polypeptide. One may also produce a fragmentby enzymatically cleaving either a recombinant polypeptide or anisolated naturally-occurring polypeptide. Methods of producingpolypeptide fragments are well-known in the art. See, e.g., Sambrook(1989), supra; Sambrook (2001), supra; Ausubel (1992), supra; andAusubel (1999), supra. In one embodiment, a polypeptide comprising onlya fragment of polypeptide of the invention, preferably a BSP, may beproduced by chemical or enzymatic cleavage of a polypeptide. In apreferred embodiment, a polypeptide fragment is produced by expressing anucleic acid molecule encoding a fragment of the polypeptide, preferablya BSP, in a host cell.

[0214] By “polypeptides” as used herein it is also meant to be inclusiveof mutants, fusion proteins, homologous proteins and allelic variants ofthe polypeptides specifically exemplified.

[0215] A mutant protein, or mutein, may have the same or differentproperties compared to a naturally-occurring polypeptide and comprisesat least one amino acid insertion, duplication, deletion, rearrangementor substitution compared to the amino acid sequence of a native protein.Small deletions and insertions can often be found that do not alter thefunction of the protein. In one embodiment, the mutein may or may not bebreast-specific. In a preferred embodiment, the mutein isbreast-specific. In a preferred embodiment, the mutein is a polypeptidethat comprises at least one amino acid insertion, duplication, deletion,rearrangement or substitution compared to the amino acid sequence of SEQID NO: 160 through 282. In a more preferred embodiment, the mutein isone that exhibits at least 50% sequence identity, more preferably atleast 60% sequence identity, even more preferably at least 70%, yet morepreferably at least 80% sequence identity to a BSP comprising an aminoacid sequence of SEQ ID NO: 160 through 282. In yet a more preferredembodiment, the mutein exhibits at least 85%, more preferably 90%, evenmore preferably 95% or 96%, and yet more preferably at least 97%, 98%,99% or 99.5% sequence identity to a BSP comprising an amino acidsequence of SEQ ID NO: 160 through 282.

[0216] A mutein may be produced by isolation from a naturally-occurringmutant cell, tissue or organism. A mutein may be produced by isolationfrom a cell, tissue or organism that has been experimentallymutagenized. Alternatively, a mutein may be produced by chemicalmanipulation of a polypeptide, such as by altering the amino acidresidue to another amino acid residue using synthetic or semi-syntheticchemical techniques. In a preferred embodiment, a mutein may be producedfrom a host cell comprising an altered nucleic acid molecule compared tothe naturally-occurring nucleic acid molecule. For instance, one mayproduce a mutein of a polypeptide by introducing one or more mutationsinto a nucleic acid sequence of the invention and then expressing itrecombinantly. These mutations may be targeted, in which particularencoded amino acids are altered, or may be untargeted, in which randomcncodcd amino acids within the polypeptide are altered. Muteins withrandom amino acid alterations can be screened for a particularbiological activity or property, particularly whether the polypeptide isbreast-specific, as described below. Multiple random mutations can beintroduced into the gene by methods well-known to the art, e.g., byerror-prone PCR, shuffling, oligonucleotide-directed mutagenesis,assembly PCR, sexual PCR mutagenesis, in vivo mutagenesis, cassettemutagenesis, recursive ensemble mutagenesis, exponential ensemblemutagenesis and site-specific mutagenesis. Methods of producing muteinswith targeted or random amino acid alterations are well-known in theart. See, e.g., Sambrook (1989), supra; Sambrook (2001), supra; Ausubel(1992), supra; and Ausubel (1999), U.S. Pat. No. 5,223,408, and thereferences discussed supra, each herein incorporated by reference.

[0217] By “polypeptide” as used herein it is also meant to be inclusiveof polypeptides homologous to those polypeptides exemplified herein. Ina preferred embodiment, the polypeptide is homologous to a BSP. In aneven more preferred embodiment, the polypeptide is homologous to a BSPselected from the group having an amino acid sequence of SEQ ID NO: 160through 282. In a preferred embodiment, the homologous polypeptide isone that exhibits significant sequence identity to a BSP. In a morepreferred embodiment, the polypeptide is one that exhibits significantsequence identity to an comprising an amino acid sequence of SEQ ID NO:160 through 282. In an even more preferred embodiment, the homologouspolypeptide is one that exhibits at least 50% sequence identity, morepreferably at least 60% sequence identity, even more preferably at least70%, yet more preferably at least 80% sequence identity to a BSPcomprising an amino acid sequence of SEQ ID NO: 160 through 282. In ayet more preferred embodiment, the homologous polypeptide is one thatexhibits at least 85%, more preferably 90%, even more preferably 95% or96%, and yet more preferably at least 97% or 98% sequence identity to aBSP comprising an amino acid sequence of SEQ ID NO: 160 through 282. Inanother preferred embodiment, the homologous polypeptide is one thatexhibits at least 99%, more preferably 99.5%, even more preferably99.6%, 99.7%, 99.8% or 99.9% sequence identity to a BSP comprising anamino acid sequence of SEQ ID NO: 160 through 282. In a preferredembodiment, the amino acid substitutions are conservative amino acidsubstitutions as discussed above.

[0218] In another embodiment, the homologous polypeptide is one that isencoded by a nucleic acid molecule that selectively hybridizes to aBSNA. In a preferred embodiment, the homologous polypeptide is encodedby a nucleic acid molecule that hybridizes to a BSNA under lowstringency, moderate stringency or high stringency conditions, asdefined herein. In a more preferred embodiment, the BSNA is selectedfrom the group consisting of SEQ ID NO: 1 through 159. In anotherpreferred embodiment, the homologous polypeptide is encoded by a nucleicacid molecule that hybridizes to a nucleic acid molecule that encodes aBSP under low stringency, moderate stringency or high stringencyconditions, as defined herein. In a more preferred embodiment, the BSPis selected from the group consisting of SEQ ID NO: 160 through 282.

[0219] The homologous polypeptide may be a naturally-occurring one thatis derived from another species, especially one derived from anotherprimate, such as chimpanzee, gorilla, rhesus macaque, baboon or gorilla,wherein the homologous polypeptide comprises an amino acid sequence thatexhibits significant sequence identity to that of SEQ ID NO: 160 through282. The homologous polypeptide may also be a naturally-occurringpolypeptide from a human, when the BSP is a member of a family ofpolypeptides. The homologous polypeptide may also be anaturally-occurring polypeptide derived from a non-primate, mammalianspecies, including without limitation, domesticated species, e.g., dog,cat, mouse, rat, rabbit, guinea pig, hamster, cow, horse, goat or pig.The homologous polypeptide may also be a naturally-occurring polypeptidederived from a non-mammalian species, such as birds or reptiles. Thenaturally-occurring homologous protein may be isolated directly fromhumans or other species. Alternatively, the nucleic acid moleculeencoding the naturally-occurring homologous polypeptide may be isolatedand used to express the homologous polypeptide recombinantly. In anotherembodiment, the homologous polypeptide may be one that is experimentallyproduced by random mutation of a nucleic acid molecule and subsequentexpression of the nucleic acid molecule. In another embodiment, thehomologous polypeptide may be one that is experimentally produced bydirected mutation of one or more codons to alter the encoded amino acidof a BSP. Further, the homologous protein may or may not encodepolypeptide that is a BSP. However, in a preferred embodiment, thehomologous polypeptide encodes a polypeptide that is a BSP.

[0220] Relatedness of proteins can also be characterized using a secondfunctional test, the ability of a first protein competitively to inhibitthe binding of a second protein to an antibody. It is, therefore,another aspect of the present invention to provide isolated proteins notonly identical in sequence to those described with particularity herein,but also to provide isolated proteins (“cross-reactive proteins”) thatcompetitively inhibit the binding of antibodies to all or to a portionof various of the isolated polypeptides of the present invention. Suchcompetitive inhibition can readily be determined using immunoassayswell-known in the art.

[0221] As discussed above, single nucleotide polymorphisms (SNPs) occurfrequently in eukaryotic genomes, and the sequence determined from oneindividual of a species may differ from other allelic forms presentwithin the population. Thus, by “polypeptide” as used herein it is alsomeant to be inclusive of polypeptides encoded by an allelic variant of anucleic acid molecule encoding a BSP. In a preferred embodiment, thepolypeptide is encoded by an allelic variant of a gene that encodes apolypeptide having the amino acid sequence selected from the groupconsisting of SEQ ID NO: 160 through 282. In a yet more preferredembodiment, the polypeptide is encoded by an allelic variant of a genethat has the nucleic acid sequence selected from the group consisting ofSEQ ID NO: 1 through 159.

[0222] In another embodiment, the invention provides polypeptides whichcomprise derivatives of a polypeptide encoded by a nucleic acid moleculeaccording to the instant invention. In a preferred embodiment, thepolypeptide is a BSP. In a preferred embodiment, the polypeptide has anamino acid sequence selected from the group consisting of SEQ ID NO: 160through 282, or is a mutein, allelic variant, homologous protein orfragment thereof. In a preferred embodiment, the derivative has beenacetylated, carboxylated, phosphorylated, glycosylated or ubiquitinated.In another preferred embodiment, the derivative has been labeled with,e.g., radioactive isotopes such as ¹²⁵I, ³²P, ³⁵S, and ³H. In anotherpreferred embodiment, the derivative has been labeled with fluorophores,chemiluminescent agents, enzymes, and antiligands that can serve asspecific binding pair members for a labeled ligand.

[0223] Polypeptide modifications are well-known to those of skill andhave been described in great detail in the scientific literature.Several particularly common modifications, glycosylation, lipidattachment, sulfation, gamma-carboxylation of glutamic acid residues,hydroxylation and ADP-ribosylation, for instance, are described in mostbasic texts, such as, for instance Creighton, Protein Structure andMolecular Properties, 2nd ed., W. H. Freeman and Company (1993). Manydetailed reviews are available on this subject, such as, for example,those provided by Wold, in Johnson (ed.), Posttranslational CovalentModification of Proteins, pgs. 1-12, Academic Press (1983); Seifter etal., Meth. Enzymol. 182: 626-646 (1990) and Rattan et al., Ann. N.Y.Acad. Sci. 663: 48-62 (1992).

[0224] It will be appreciated, as is well-known and as noted above, thatpolypeptides are not always entirely linear. For instance, polypeptidesmay be branched as a result of ubiquitination, and they may be circular,with or without branching, generally as a result of posttranslationevents, including natural processing event and events brought about byhuman manipulation which do not occur naturally. Circular, branched andbranched circular polypeptides may be synthesized by non-translationnatural process and by entirely synthetic methods, as well.Modifications can occur anywhere in a polypeptide, including the peptidebackbone, the amino acid side-chains and the amino or carboxyl termini.In fact, blockage of the amino or carboxyl group in a polypeptide, orboth, by a covalent modification, is common in naturally occurring andsynthetic polypeptides and such modifications may be present inpolypeptides of the present invention, as well. For instance, the aminoterminal residue of polypeptides made in E. coli, prior to proteolyticprocessing, almost invariably will be N-formylmethionine.

[0225] Useful post-synthetic (and post-translational) modificationsinclude conjugation to detectable labels, such as fluorophores. A widevariety of amine-reactive and thiol-reactive fluorophore derivativeshave been synthesized that react under nondenaturing conditions withN-terminal amino groups and epsilon amino groups of lysine residues, onthe one hand, and with free thiol groups of cysteine residues, on theother.

[0226] Kits are available commercially that permit conjugation ofproteins to a variety of amine-reactive or thiol-reactive fluorophores:Molecular Probes, Inc. (Eugene, Oreg., USA), e.g., offers kits forconjugating proteins to Alexa Fluor 350, Alexa Fluor 430,Fluorescein-EX, Alexa Fluor 488, Oregon Green 488, Alexa Fluor 532,Alexa Fluor 546, Alexa Fluor 546, Alexa Fluor 568, Alexa Fluor 594, andTexas Red-X.

[0227] A wide variety of other amine-reactive and thiol-reactivefluorophores are available commercially (Molecular Probes, Inc., Eugene,Oreg., USA), including Alexa Fluor® 350, Alexa Fluor® 488, Alexa Fluor®532, Alexa Fluor® 546, Alexa Fluor® 568, Alexa Fluor® 594, Alexa Fluor®647 (monoclonal antibody labeling kits available from Molecular Probes,Inc., Eugene, Oreg., USA), BODIPY dyes, such as BODIPY 493/503, BODIPYFL, BODIPY R6G, BODIPY 530/550, BODIPY TMR, BODIPY 558/568, BODIPY558/568, BODIPY 564/570, BODIPY 576/589, BODIPY 581/591, BODIPY TR,BODIPY 630/650, BODIPY 650/665, Cascade Blue, Cascade Yellow, Dansyl,lissamine rhodamine B, Marina Blue, Oregon Green 488, Oregon Green 514,Pacific Blue, rhodamine 6G, rhodamine green, rhodamine red,tetramethylrhodamine, Texas Red (available from Molecular Probes, Inc.,Eugene, Oreg., USA).

[0228] The polypeptides of the present invention can also be conjugatedto fluorophores, other proteins, and other macromolecules, usingbifunctional linking reagents. Common homobifunctional reagents include,e.g., APG, AEDP, BASED, BMB, BMDB, BMH, BMOE, BM[PEO]3, BM[PEO]4, BS3,BSOCOES, DFDNB, DMA, DMP, DMS, DPDPB, DSG, DSP (Lomant's Reagent), DSS,DST, DTBP, DTME, DTSSP, EGS, HBVS, Sulfo-BSOCOES, Sulfo-DST, Sulfo-EGS(all available from Pierce, Rockford, Ill., USA); commonheterobifunctional cross-linkers include ABH, AMAS, ANB-NOS, APDP, ASBA,BMPA, BMPH, BMPS, EDC, EMCA, EMCH, EMCS, KMUA, KMUH, GMBS, LC-SMCC,LC-SPDP, MBS, M2C2H, MPBH, MSA, NHS-ASA, PDPH, PMPI, SADP, SAED, SAND,SANPAH, SASD, SATP, SBAP, SFAD, SIA, SIAB, SMCC, SMPB, SMPH, SMPT, SPDP,Sulfo-EMCS, Sulfo-GMBS, Sulfo-HSAB, Sulfo-KMUS, Sulfo-LC-SPDP,Sulfo-MBS, Sulfo-NHS-LC-ASA, Sulfo-SADP, Sulfo-SANPAH, Sulfo-SIAB,Sulfo-SMCC, Sulfo-SMPB, Sulfo-LC-SMPT, SVSB, TFCS (all available Pierce,Rockford, Ill., USA).

[0229] The polypeptides, fragments, and fusion proteins of the presentinvention can be conjugated, using such cross-linking reagents, tofluorophores that are not amine- or thiol-reactive. Other labels thatusefully can be conjugated to the polypeptides, fragments, and fusionproteins of the present invention include radioactive labels,echosonographic contrast reagents, and MRI contrast agents.

[0230] The polypeptides, fragments, and fusion proteins of the presentinvention can also usefully be conjugated using cross-linking agents tocarrier proteins, such as KLH, bovine thyroglobulin, and even bovineserum albumin (BSA), to increase immunogenicity for raising anti-BSPantibodies.

[0231] The polypeptides, fragments, and fusion proteins of the presentinvention can also usefully be conjugated to polyethylene glycol (PEG);PEGylation increases the serum half-life of proteins administeredintravenously for replacement therapy. Delgado et al., Crit. Rev. Ther.Drug Carrier Syst. 9(3-4): 249-304 (1992); Scott et al., Curr. Pharm.Des. 4(6): 423-38 (1998); DeSantis et al., Curr. Opin. Biotechnol.10(4): 324-30 (1999) incorporated herein by reference in theirentireties. PEG monomers can be attached to the protein directly orthrough a linker, with PEGylation using PEG monomers activated withtresyl chloride (2,2,2-trifluoroethanesulphonyl chloride) permittingdirect attachment under mild conditions.

[0232] In yet another embodiment, the invention provides analogs of apolypeptide encoded by a nucleic acid molecule according to the instantinvention. In a preferred embodiment, the polypeptide is a BSP. In amore preferred embodiment, the analog is derived from a polypeptidehaving part or all of the amino acid sequence of SEQ ID NO: 160 through282. In a preferred embodiment, the analog is one that comprises one ormore substitutions of non-natural amino acids or non-nativeinter-residue bonds compared to the naturally-occurring polypeptide. Ingeneral, the non-peptide analog is structurally similar to a BSP, butone or more peptide linkages is replaced by a linkagc selected from thegroup consisting of ——CH₂NH——, ——CH₂S——, ——CH₂—CH₂——, ——CH═CH—— (cis andtrans), ——COCH₂——, ——CH(OH)CH₂—— and —CH₂SO——. In another embodiment,the non-peptide analog comprises substitution of one or more amino acidsof a BSP with a D-amino acid of the same type or other non-natural aminoacid in order to generate more stable peptides. D-amino acids canreadily be incorporated during chemical peptide synthesis: peptidesassembled from D-amino acids are more resistant to proteolytic attack;incorporation of D-amino acids can also be used to confer specificthree-dimensional conformations on the peptide. Other amino acidanalogues commonly added during chemical synthesis include ornithine,norleucine, phosphorylated amino acids (typically phosphoserine,phosphothreonine, phosphotyrosine), L-malonyltyrosine, anon-hydrolyzable analog of phosphotyrosine (see, e.g., Kole et al.,Biochem. Biophys. Res. Com. 209: 817-821 (1995)), and varioushalogenated phenylalanine derivatives.

[0233] Non-natural amino acids can be incorporated during solid phasechemical synthesis or by recombinant techniques, although the former istypically more common. Solid phase chemical synthesis of peptides iswell established in the art. Procedures are described, inter alia, inChan et al. (eds.), Fmoc Solid Phase Peptide Synthesis: A PracticalApproach (Practical Approach Series), Oxford Univ. Press (March 2000);Jones, Amino Acid and Peptide Synthesis (Oxford Chemistry Primers, No7), Oxford Univ. Press (1992); and Bodanszky, Principles of PeptideSynthesis (Springer Laboratory), Springer Verlag (1993); the disclosuresof which are incorporated herein by reference in their entireties.

[0234] Amino acid analogues having detectable labels are also usefullyincorporated during synthesis to provide derivatives and analogs.Biotin, for example can be added usingbiotinoyl-(9-fluorenylmethoxycarbonyl)-L-lysine (FMOC biocytin)(Molecular Probes, Eugene, Oreg., USA). Biotin can also be addedenzymatically by incorporation into a fusion protein of a E. coli BirAsubstrate peptide. The FMOC and tBOC derivatives of dabcyl-L-lysine(Molecular Probes, Inc., Eugene, Oreg., USA) can be used to incorporatethe dabcyl chromophore at selected sites in the peptide sequence duringsynthesis. The aminonaphthalene derivative EDANS, the most commonfluorophore for pairing with the dabcyl quencher in fluorescenceresonance energy transfer (FRET) systems, can be introduced duringautomated synthesis of peptides by using EDANS-FMOC-L-glutamic acid orthe corresponding tBOC derivative (both from Molecular Probes, Inc.,Eugene, Oreg., USA). Tetramethylrhodamine fluorophores can beincorporated during automated FMOC synthesis of peptides using(FMOC)-TMR-L-lysine (Molecular Probes, Inc. Eugene, Oreg., USA).

[0235] Other useful amino acid analogues that can be incorporated duringchemical synthesis include aspartic acid, glutamic acid, lysine, andtyrosine analogues having allyl side-chain protection (AppliedBiosystems, Inc., Foster City, Calif., USA); the allyl side chainpermits synthesis of cyclic, branched-chain, sulfonated, glycosylated,and phosphorylated peptides.

[0236] A large number of other FMOC-protected non-natural amino acidanalogues capable of incorporation during chemical synthesis areavailable commercially, including, e.g., Fmoc-2-aminobicyclo[2.2.1]heptane-2-carboxylic acid, Fmoc-3-endo-aminobicyclo[2.2.1]heptane-2-endo-carboxylic acid, Fmoc-3-exo-aminobicyclo[2.2.1]heptane-2-exo-carboxylic acid, Fmoc-3-endo-aminobicyclo[2.2. 1]hept-5-ene-2-endo-carboxylic acid, Fmoc-3-exo-amino-bicyclo[2.2. 1]hept-5-ene-2-exo-carboxylic acid,Fmoc-cis-2-amino-1-cyclohexanecarboxylic acid,Fmoc-trans-2-amino-1-cyclohexanecarboxylic acid,Fmoc-1-amino-1-cyclopentanecarboxylic acid,Fmoc-cis-2-amino-1-cyclopentanecarboxylic acid,Fmoc-1-amino-1-cyclopropanecarboxylic acid,Fmoc-D-2-amino-4-(ethylthio)butyric acid,Fmoc-L-2-amino-4-(ethylthio)butyric acid, Fmoc-L-buthionine,Fmoc-S-methyl-L-Cysteine, Fmoc-2-aminobenzoic acid (anthranillic acid),Fmoc-3-aminobenzoic acid, Fmoc-4-aminobenzoic acid,Fmoc-2-aminobenzophenone-2′-carboxylic acid,Fmoc-N-(4-aminobenzoyl)-β-alanine, Fmoc-2-amino-4,5-dimethoxybenzoicacid, Fmoc-4-aminohippuric acid, Fmoc-2-amino-3-hydroxybenzoic acid,Fmoc-2-amino-5-hydroxybenzoic acid, Fmoc-3-amino-4-hydroxybenzoic acid,Fmoc-4-amino-3-hydroxybenzoic acid, Fmoc-4-amino-2-hydroxybenzoic acid,Fmoc-5-amino-2-hydroxybenzoic acid, Fmoc-2-amino-3-methoxybenzoic acid,Fmoc-4-amino-3-methoxybenzoic acid, Fmoc-2-amino-3-methylbenzoic acid,Fmoc-2-amino-5-methylbenzoic acid, Fmoc-2-amino-6-methylbenzoic acid,Fmoc-3-amino-2-methylbenzoic acid, Fmoc-3-amino-4-methylbenzoic acid,Fmoc-4-amino-3-methylbenzoic acid, Fmoc-3-amino-2-naphtoic acid,Fmoc-D,L-3-amino-3-phenylpropionic acid, Fmoc-L-Methyldopa,Fmoc-2-amino-4,6-dimethyl-3-pyridinecarboxylic acid,Fmoc-D,L-amino-2-thiophenacetic acid, Fmoc-4-(carboxymethyl)piperazine,Fmoc-4-carboxypiperazine, Fmoc-4-(carboxymethyl)homopiperazine,Fmoc-4-phenyl-4-piperidinecarboxylic acid,Fmoc-L-1,2,3,4-tetrahydronorharman-3-carboxylic acid,Fmoc-L-thiazolidine-4-carboxylic acid, all available from The PeptideLaboratory (Richmond, Calif., USA).

[0237] Non-natural residues can also be added biosynthetically byengineering a suppressor tRNA, typically one that recognizes the UAGstop codon, by chemical aminoacylation with the desired unnatural aminoacid. Conventional site-directed mutagenesis is used to introduce thechosen stop codon UAG at the site of interest in the protein gene. Whenthe acylated suppressor tRNA and the mutant gene are combined in an invitro transcription/translation system, the unnatural amino acid isincorporated in response to the UAG codon to give a protein containingthat amino acid at the specified position. Liu et al., Proc. Natl Acad.Sci. USA 96(9): 4780-5 (1999); Wang et al., Science 292(5516): 498-500(2001).

[0238] Fusion Proteins

[0239] The present invention further provides fusions of each of thepolypeptides and fragments of the present invention to heterologouspolypeptides. In a preferred embodiment, the polypeptide is a BSP. In amore preferred embodiment, the polypeptide that is fused to theheterologous polypeptide comprises part or all of the amino acidsequence of SEQ ID NO: 160 through 282, or is a mutein, homologouspolypeptide, analog or derivative thereof. In an even more preferredembodiment, the nucleic acid molecule encoding the fusion proteincomprises all or part of the nucleic acid sequence of SEQ ID NO: 1through 159, or comprises all or part of a nucleic acid sequence thatselectively hybridizes or is homologous to a nucleic acid moleculecomprising a nucleic acid sequence of SEQ ID NO: 1 through 159.

[0240] The fusion proteins of the present invention will include atleast one fragment of the protein of the present invention, whichfragment is at least 6, typically at least 8, often at least 15, andusefully at least 16, 17, 18, 19, or 20 amino acids long. The fragmentof the protein of the present to be included in the fusion can usefullybe at least 25 amino acids long, at least 50 amino acids long, and canbe at least 75, 100, or even 150 amino acids long. Fusions that includethe entirety of the proteins of the present invention have particularutility.

[0241] The heterologous polypeptide included within the fusion proteinof the present invention is at least 6 amino acids in length, often atleast 8 amino acids in length, and usefully at least 15, 20, and 25amino acids in length. Fusions that include larger polypeptides, such asthe IgG Fe region, and even entire proteins (such as GFPchromophore-containing proteins) are particular useful.

[0242] As described above in the description of vectors and expressionvectors of the present invention, which discussion is incorporated hereby reference in its entirety, heterologous polypeptides to be includedin the fusion proteins of the present invention can usefully includethose designed to facilitate purification and/or visualization ofrecombinantly-expressed proteins. See, e.g., Ausubel, Chapter 16,(1992), supra. Although purification tags can also be incorporated intofusions that are chemically synthesized, chemical synthesis typicallyprovides sufficient purity that further purification by HPLC suffices;however, visualization tags as above described retain their utility evenwhen the protein is produced by chemical synthesis, and when so includedrender the fusion proteins of the present invention useful as directlydetectable markers of the presence of a polypeptide of the invention.

[0243] As also discussed above, heterologous polypeptides to be includedin the fusion proteins of the present invention can usefully includethose that facilitate secretion of recombinantly expressed proteins—intothe periplasmic space or extracellular milieu for prokaryotic hosts,into the culture medium for eukaryotic cells—through incorporation ofsecretion signals and/or leader sequences. For example, a His⁶ taggedprotein can be purified on a Ni affinity column and a GST fusion proteincan be purified on a glutathione affinity column. Similarly, a fusionprotein comprising the Fc domain of IgG can be purified on a Protein Aor Protein G column and a fusion protein comprising an epitope tag suchas myc can be purified using an immunoaffinity column containing ananti-c-myc antibody. It is preferable that the epitope tag be separatedfrom the protein encoded by the essential gene by an enzymatic cleavagesite that can be cleaved after purification. See also the discussion ofnucleic acid molecules encoding fusion proteins that may be expressed onthe surface of a cell.

[0244] Other useful protein fusions of the present invention includethose that permit use of the protein of the present invention as bait ina yeast two-hybrid system. See Bartel et al. (eds.), The YeastTwo-Hybrid System, Oxford University Press (1997); Zhu et al., YeastHybrid Technologies, Eaton Publishing (2000); Fields et al., TrendsGenet. 10(8): 286-92 (1994); Mendelsohn et al., Curr. Opin. Biotechnol.5(5): 482-6 (1994); Luban et al., Curr. Opin. Biotechnol. 6(1): 59-64(1995); Allen et al., Trends Biochem. Sci. 20(12): 511-6 (1995); Drees,Curr. Opin. Chem. Biol. 3(1): 64-70 (1999); Topcu et al., Pharm. Res.17(9): 1049-55 (2000); Fashena et al., Gene 250(1-2): 1-14 (2000);;Colas et al., (1996) Genetic selection of peptide aptamers thatrecognize and inhibit cyclin-dependent kinase 2. Nature 380, 548-550;Norman, T. et al., (1999) Genetic selection of peptide inhibitors ofbiological pathways. Science 285, 591-595, Fabbrizio et al., (1999)Inhibition of mammalian cell proliferation by genetically selectedpeptide aptamers that functionally antagonize E2F activity. Oncogene 18,4357-4363; Xu et al., (1997) Cells that register logical relationshipsamong proteins. Proc Natl Acad Sci USA. 94, 12473-12478; Yang, et al.,(1995) Protein-peptide interactions analyzed with the yeast two-hybridsystem. Nuc. Acids Res. 23, 1152-1156; Kolonin et al., (1998) Targetingcyclin-dependent kinases in Drosophila with peptide aptamers. Proc NatlAcad Sci USA 95, 14266-14271; Cohen et al., (1998) An artificialcell-cycle inhibitor isolated from a combinatorial library. Proc NatlAcad Sci USA 95, 14272-14277; Uetz, P.; Giot, L.; al, e.; Fields, S.;Rothberg, J. M. (2000) A comprehensive analysis of protein-proteininteractions in Saccharomyces cerevisiae. Nature 403, 623-627; Ito, etal., (2001) A comprehensive two-hybrid analysis to explore the yeastprotein interactome. Proc Natl Acad Sci USA 98, 4569-4574, thedisclosures of which are incorporated herein by reference in theirentireties. Typically, such fusion is to either E. coli LexA or yeastGAL4 DNA binding domains. Related bait plasmids are available thatexpress the bait fused to a nuclear localization signal.

[0245] Other useful fusion proteins include those that permit display ofthe encoded protein on the surface of a phage or cell, fusions tointrinsically fluorescent proteins, such as green fluorescent protein(GFP), and fusions to the IgG Fc region, as described above, whichdiscussion is incorporated here by reference in its entirety.

[0246] The polypeptides and fragments of the present invention can alsousefully be fused to protein toxins, such as Pseudomonas exotoxin A,diphtheria toxin, shiga toxin A, anthrax toxin lethal factor, ricin, inorder to effect ablation of cells that bind or take up the proteins ofthe present invention.

[0247] Fusion partners include, inter alia, myc, hcmagglutinin (HA),GST, immunoglobulins, β-galactosidase, biotin trpE, protein A,β-lactamase, -amylase, maltose binding protein, alcohol dehydrogenase,polyhistidine (for example, six histidine at the amino and/or carboxylterminus of the polypeptide), lacZ, green fluorescent protein (GFP),yeast_mating factor, GAL4 transcription activation or DNA bindingdomain, luciferase, and serum proteins such as ovalbumin, albumin andthe constant domain of IgG. See, e.g., Ausubel (1992), supra and Ausubel(1999), supra. Fusion proteins may also contain sites for specificenzymatic cleavage, such as a site that is recognized by enzymes such asFactor XIII, trypsin, pepsin, or any other enzyme known in the art.Fusion proteins will typically be made by either recombinant nucleicacid methods, as described above, chemically synthesized usingtechniques well-known in the art (e.g., a Merrifield synthesis), orproduced by chemical cross-linking.

[0248] Another advantage of fusion proteins is that the epitope tag canbe used to bind the fusion protein to a plate or column through anaffinity linkage for screening binding proteins or other molecules thatbind to the BSP.

[0249] As further described below, the isolated polypeptides, muteins,fusion proteins, homologous proteins or allelic variants of the presentinvention can readily be used as specific immunogens to raise antibodiesthat specifically recognize BSPs, their allelic variants and homologues.The antibodies, in turn, can be used, inter alia, specifically to assayfor the polypeptides of the present invention, particularly BSPs, e.g.by ELISA for detection of protein fluid samples, such as serum, byimmunohistochemistry or laser scanning cytometry, for detection ofprotein in tissue samples, or by flow cytometry, for detection ofintracellular protein in cell suspensions, for specificantibody-mediated isolation and/or purification of BSPs, as for exampleby immunoprecipitation, and for use as specific agonists or antagonistsof BSPs.

[0250] One may determine whether polypeptides including muteins, fusionproteins, homologous proteins or allelic variants are functional bymethods known in the art. For instance, residues that are tolerant ofchange while retaining function can be identified by altering theprotein at known residues using methods known in the art, such asalanine scanning mutagenesis, Cunningham et al., Science 244(4908):1081-5 (1989); transposon linker scanning mutagenesis, Chen et al., Gene263(1-2): 39-48 (2001); combinations of homolog- and alanine-scanningmutagenesis, Jin et al., J. Mol. Biol. 226(3): 851-65 (1992);combinatorial alaninc scanning, Weiss et al., Proc. Natl. Acad. Sci USA97(16): 8950-4 (2000), followed by functional assay. Transposon linkerscanning kits are available commercially (New England Biolabs, Beverly,Mass., USA, catalog. no. E7-102S; EZ::TN™ In-Frame Linker Insertion Kit,catalogue no. EZI04KN, Epicentre Technologies Corporation, Madison,Wis., USA).

[0251] Purification of the polypeptides including fragments, homologouspolypeptides, muteins, analogs, derivatives and fusion proteins iswell-known and within the skill of one having ordinary skill in the art.See, e.g., Scopes, Protein Purification, 2d ed. (1987). Purification ofrecombinantly expressed polypeptides is described above. Purification ofchemically-synthesized peptides can readily be effected, e.g., by HPLC.

[0252] Accordingly, it is an aspect of the present invention to providethe isolated proteins of the present invention in pure or substantiallypure form in the presence of absence of a stabilizing agent. Stabilizingagents include both proteinaceous or non-proteinaceous material and arewell-known in the art. Stabilizing agents, such as albumin andpolyethylene glycol (PEG) are known and are commercially available.

[0253] Although high levels of purity are preferred when the isolatedproteins of the present invention are used as therapeutic agents, suchas in vaccines and as replacement therapy, the isolated proteins of thepresent invention are also useful at lower purity. For example,partially purified proteins of the present invention can be used asimmunogens to raise antibodies in laboratory animals.

[0254] In preferred embodiments, the purified and substantially purifiedproteins of the present invention are in compositions that lackdetectable ampholytes, acrylamide monomers, bis-acrylamide monomers, andpolyacrylamide.

[0255] The polypeptides, fragments, analogs, derivatives and fusions ofthe present invention can usefully be attached to a substrate. Thesubstrate can be porous or solid, planar or non-planar; the bond can becovalent or noncovalent.

[0256] For example, the polypeptides, fragments, analogs, derivativesand fusions of the present invention can usefully be bound to a poroussubstrate, commonly a membrane, typically comprising nitrocellulose,polyvinylidene fluoride (PVDF), or cationically derivatized, hydrophilicPVDF; so bound, the proteins, fragments, and fusions of the presentinvention can be used to detect and quantify antibodies, e.g. in serum,that bind specifically to the immobilized protein of the presentinvention.

[0257] As another example, the polypeptides, fragments, analogs,derivatives and fusions of the present invention can usefully be boundto a substantially nonporous substrate, such as plastic, to detect andquantify antibodies, e.g. in serum, that bind specifically to theimmobilized protein of the present invention. Such plastics includepolymethylacrylic, polyethylene, polypropylene, polyacrylate,polymethylmethacrylate, polyvinylchloride, polytetrafluoroethylene,polystyrene, polycarbonate, polyacetal, polysulfone, celluloseacetate,cellulosenitrate, nitrocellulose, or mixtures thereof; when the assay isperformed in a standard microtiter dish, the plastic is typicallypolystyrene.

[0258] The polypeptides, fragments, analogs, derivatives and fusions ofthe present invention can also be attached to a substrate suitable foruse as a surface enhanced laser desorption ionization source; soattached, the protein, fragment, or fusion of the present invention isuseful for binding and then detecting secondary proteins that bind withsufficient affinity or avidity to the surface-bound protein to indicatebiologic interaction there between. The proteins, fragments, and fusionsof the present invention can also be attached to a substrate suitablefor usc in surface plasmon resonance detection; so attached, theprotein, fragment, or fusion of the present invention is useful forbinding and then detecting secondary proteins that bind with sufficientaffinity or avidity to the surface-bound protein to indicate biologicalinteraction there between.

[0259] Antibodies

[0260] In another aspect, the invention provides antibodies, includingfragments and derivatives thereof, that bind specifically topolypeptides encoded by the nucleic acid molecules of the invention, aswell as antibodies that bind to fragments, muteins, derivatives andanalogs of the polypeptides. In a preferred embodiment, the antibodiesare specific for a polypeptide that is a BSP, or a fragment, mutein,derivative, analog or fusion protein thereof. In a more preferredembodiment, the antibodies are specific for a polypeptide that comprisesSEQ ID NO: 160 through 282, or a fragment, mutein, derivative, analog orfusion protein thereof.

[0261] The antibodies of the present invention can be specific forlinear epitopes, discontinuous epitopes, or conformational epitopes ofsuch proteins or protein fragments, either as present on the protein inits native conformation or, in some cases, as present on the proteins asdenatured, as, e.g., by solubilization in SDS. New epitopes may be alsodue to a difference in post translational modifications (PTMs) indisease versus normal tissue. For example, a particular site on a BSPmay be glycosylated in cancerous cells, but not glycosylated in normalcells or visa versa. In addition, alternative splice forms of a BSP maybe indicative of cancer. Differential degradation of the C or N-terminusof a BSP may also be a marker or target for anticancer therapy. Forexample, a BSP may be N-terminal degraded in cancer cells exposing newepitopes to which antibodies may selectively bind for diagnostic ortherapeutic uses.

[0262] As is well-known in the art, the degree to which an antibody candiscriminate as among molecular species in a mixture will depend, inpart, upon the conformational relatedness of the species in the mixture;typically, the antibodies of the present invention will discriminateover adventitious binding to non-BSP polypeptides by at least 2-fold,more typically by at least 5-fold, typically by more than 10-fold,25-fold, 50-fold, 75-fold, and often by more than 100-fold, and onoccasion by more than 500-fold or 1000-fold. When used to detect theproteins or protein fragments of the present invention, the antibody ofthe present invention is sufficiently specific when it can be used todetermine the presence of the protein of the present invention insamples derived from human breast.

[0263] Typically, the affinity or avidity of an antibody (or antibodymultimer, as in the case of an IgM pentamer) of the present inventionfor a protein or protein fragment of the present invention will be atleast about 1×10⁻⁶ molar (M), typically at least about 5×10⁻⁷ M, 1×10⁻⁷M, with affinities and avidities of at least 1×10⁻⁸ M, 5×10⁻⁹ M, 1×10⁻¹⁰M and up to 1×10⁻¹³ M proving especially useful.

[0264] The antibodies of the present invention can benaturally-occurring forms, such as IgG, IgM, IgD, IgE, IgY, and IgA,from any avian, reptilian, or mammalian species.

[0265] Human antibodies can, but will infrequently, be drawn directlyfrom human donors or human cells. In this case, antibodies to theproteins of the present invention will typically have resulted fromfortuitous immunization, such as autoimmune immunization, with theprotein or protein fragments of the present invention. Such antibodieswill typically, but will not invariably, be polyclonal. In addition,individual polyclonal antibodies may be isolated and cloned to generatemonoclonals.

[0266] Human antibodies are more frequently obtained using transgenicanimals that express human immunoglobulin genes, which transgenicanimals can be affirmatively immunized with the protein immunogen of thepresent invention. Human Ig-transgenic mice capable of producing humanantibodies and methods of producing human antibodies therefrom uponspecific immunization are described, inter alia, in U.S. Pat. Nos.6,162,963; 6,150,584; 6,114,598; 6,075,181; 5,939,598; 5,877,397;5,874,299; 5,814,318; 5,789,650; 5,770,429; 5,661,016; 5,633,425;5,625,126; 5,569,825; 5,545,807; 5,545,806, and 5,591,669, thedisclosures of which are incorporated herein by reference in theirentireties. Such antibodies are typically monoclonal, and are typicallyproduced using techniques developed for production of murine antibodies.

[0267] Human antibodies are particularly useful, and often preferred,when the antibodies of the present invention are to be administered tohuman beings as in vivo diagnostic or therapeutic agents, sincerecipient immune response to the administered antibody will often besubstantially less than that occasioned by administration of an antibodyderived from another species, such as mouse.

[0268] IgG, IgM, IgD, IgE, IgY, and IgA antibodies of the presentinvention can also be obtained from other species, including mammalssuch as rodents (typically mouse, but also rat, guinea pig, and hamster)lagomorphs, typically rabbits, and also larger mammals, such as sheep,goats, cows, and horses, and other egg laying birds or reptiles such aschickens or alligators. For example, avian antibodies may be generatedusing techniques described in WO 00/29444, published May 25, 2000, thecontents of which are hereby incorporated in their entirety. In suchcases, as with the transgenic human-antibody-producing non-humanmammals, fortuitous immunization is not required, and the non-humanmammal is typically affirmatively immunized, according to standardimmunization protocols, with the protein or protein fragment of thepresent invention.

[0269] As discussed above, virtually all fragments of 8 or morecontiguous amino acids of the proteins of the present invention can beused effectively as immunogens when conjugated to a carrier, typically aprotein such as bovine thyroglobulin, keyhole limpet hemocyanin, orbovine serum albumin, conveniently using a bifunctional linker such asthose described elsewhere above, which discussion is incorporated byreference here.

[0270] Immunogenicity can also be conferred by fusion of the polypeptideand fragments of the present invention to other moieties. For example,peptides of the present invention can be produced by solid phasesynthesis on a branched polylysine core matrix; these multiple antigenicpeptides (MAPs) provide high purity, increased avidity, accuratechemical definition and improved safety in vaccine development. Tam etal., Proc. Natl. Acad. Sci. USA 85: 5409-5413 (1988); Posnett et al., J.Biol. Chem. 263: 1719-1725 (1988).

[0271] Protocols for immunizing non-human mammals or avian species arewell-established in the art. See Harlow et al. (eds.), Using Antibodies:A Laboratory Manual, Cold Spring Harbor Laboratory (1998); Coligan etal. (eds.), Current Protocols in Immunology, John Wiley & Sons, Inc.(2001); Zola, Monoclonal Antibodies: Preparation and Use of MonoclonalAntibodies and Engineered Antibody Derivatives (Basics: From Backgroundto Bench), Springer Verlag (2000); Gross M, Speck J.Dtsch. Tierarztl.Wochenschr. 103: 417-422 (1996), the disclosures of which areincorporated herein by reference. Immunization protocols often includemultiple immunizations, either with or without adjuvants such asFreund's complete adjuvant and Freund's incomplete adjuvant, and mayinclude naked DNA immunization (Moss, Semin. Immunol. 2: 317-327 (1990).

[0272] Antibodies from non-human mammals and avian species can bepolyclonal or monoclonal, with polyclonal antibodies having certainadvantages in immunohistochemical detection of the proteins of thepresent invention and monoclonal antibodies having advantages inidentifying and distinguishing particular epitopes of the proteins ofthe present invention. Antibodies from avian species may have particularadvantage in detection of the proteins of the present invention, inhuman serum or tissues (Vikinge et al., Biosens. Bioelectron. 13:1257-1262 (1998).

[0273] Following immunization, the antibodies of the present inventioncan be produced using any art-accepted technique. Such techniques arewell-known in the art, Coligan, supra; Zola, supra; Howard et al.(eds.), Basic Methods in Antibody Production and Characterization, CRCPress (2000); Harlow, supra; Davis (ed.), Monoclonal Antibody Protocols,Vol. 45, Humana Press (1995); Delves (ed.), Antibody Production:Essential Techniques, John Wiley & Son Ltd (1997); Kenney, AntibodySolution: An Antibody Methods Manual, Chapman & Hall (1997),incorporated herein by reference in their entireties, and thus need notbe detailed here.

[0274] Briefly, however, such techniques include, inter alia, productionof monoclonal antibodies by hybridomas and expression of antibodics orfragments or derivatives thereof from host cells engineered to expressimmunoglobulin genes or fragments thereof. These two methods ofproduction are not mutually exclusive: genes encoding antibodiesspecific for the proteins or protein fragments of the present inventioncan be cloned from hybridomas and thereafter expressed in other hostcells. Nor need the two necessarily be performed together: e.g., genesencoding antibodies specific for the proteins and protein fragments ofthe present invention can be cloned directly from B cells known to bespecific for the desired protein, as further described in U.S Pat. No.5,627,052, the disclosure of which is incorporated herein by referencein its entirety, or from antibody-displaying phage.

[0275] Recombinant expression in host cells is particularly useful whenfragments or derivatives of the antibodies of the present invention aredesired.

[0276] Host cells for recombinant production of either whole antibodies,antibody fragments, or antibody derivatives can be prokaryotic oreukaryotic.

[0277] Prokaryotic hosts are particularly useful for producing phagedisplayed antibodies of the present invention.

[0278] The technology of phage-displayed antibodies, in which antibodyvariable region fragments are fused, for example, to the gene IIIprotein (pIII) or gene VIII protein (pVIII) for display on the surfaceof filamentous phage, such as M13, is by now well-established. See,e.g., Sidhu, Curr. Opin. Biotechnol. 11(6): 610-6 (2000); Griffiths etal., Curr. Opin. Biotechnol. 9(1): 102-8 (1998); Hoogenboom et al.,Immunotechnology, 4(1): 1-20 (1998); Rader et al., Current Opinion inBiotechnology 8: 503-508 (1997); Aujame et al., Human Antibodies 8:155-168 (1997); Hoogenboom, Trends in Biotechnol. 15: 62-70 (1997); deKruif et al., 17: 453-455 (1996); Barbas et al., Trends in Biotechnol.14: 230-234 (1996); Winter et al., Ann. Rev. Immunol. 433-455 (1994).Techniques and protocols required to generate, propagate, screen (pan),and use the antibody fragments from such libraries have recently beencompiled. See, e.g., Barbas (2001), supra; Kay, supra; Abelson, supra,the disclosures of which are incorporated herein by reference in theirentireties.

[0279] Typically, phage-displayed antibody fragments are scFv fragmentsor Fab fragments; when desired, full length antibodies can be producedby cloning the variable regions from the displaying phage into acomplete antibody and expressing the full length antibody in a furtherprokaryotic or a cukaryotic host cell.

[0280] Eukaryotic cells are also useful for expression of theantibodies, antibody fragments, and antibody derivatives of the presentinvention.

[0281] For example, antibody fragments of the present invention can beproduced in Pichia pastoris and in Saccharomyces cerevisiae. See, e.g.,Takahashi et al., Biosci. Biotechnol. Biochem. 64(10): 2138-44 (2000);Freyre et al., J. Biotechnol. 76(2-3):1 57-63 (2000); Fischer et al.,Biotechnol. Appl. Biochem. 30 (Pt 2): 117-20 (1999); Pennell et al.,Res. Immunol. 149(6): 599-603 (1998); Eldin et al., J. Immunol. Methods.201(1): 67-75 (1997);, Frenken et al., Res. Immunol. 149(6): 589-99(1998); Shusta et al., Nature Biotechnol. 16(8): 773-7 (1998), thedisclosures of which are incorporated herein by reference in theirentireties.

[0282] Antibodies, including antibody fragments and derivatives, of thepresent invention can also be produced in insect cells. See, e.g., Li etal., Protein Expr. Purif. 21(1): 121-8 (2001); Ailor et al., Biotechnol.Bioeng. 58(2-3): 196-203 (1998); Hsu et al., Biotechnol. Prog. 13(1):96-104 (1997); Edelman et al., Immunology 91(1): 13-9 (1997); and Nesbitet al., J. Immunol. Methods 151(1-2): 201-8 (1992), the disclosures ofwhich are incorporated herein by reference in their entireties.

[0283] Antibodies and fragments and derivatives thereof of the presentinvention can also be produced in plant cells, particularly maize ortobacco, Giddings et al., Nature Biotechnol. 18(11): 1151-5 (2000);Gavilondo et al., Biotechniques 29(1): 128-38 (2000); Fischer et al., J.Biol. Regul. Homeost. Agents 14(2): 83-92 (2000); Fischer et al.,Biotechnol. Appl. Biochem. 30 (Pt 2): 113-6 (1999); Fischer et al.,Biol. Chem. 380(7-8): 825-39 (1999); Russell, Curr. Top. Microbiol.Immunol. 240: 119-38 (1999); and Ma et al., Plant Physiol. 109(2): 341-6(1995), the disclosures of which are incorporated herein by reference intheir entireties.

[0284] Antibodies, including antibody fragments and derivatives, of thepresent invention can also be produced in transgenic, non-human,mammalian milk. See, e.g. Pollock et al., J. Immunol Methods. 231:147-57 (1999); Young et al., Res. Immunol. 149: 609-10 (1998); Limontaet al., Immunotechnology 1: 107-13 (1995), the disclosures of which areincorporated herein by reference in their entireties.

[0285] Mammalian cells useful for recombinant expression of antibodies,antibody fragments, and antibody derivatives of the present inventioninclude CHO cells, COS cells, 293 cells, and mycloma cells.

[0286] Verma et al., J. Immunol. Methods 216(1-2):165-81 (1998), hereinincorporated by reference, review and compare bacterial, yeast, insectand mammalian expression systems for expression of antibodies.

[0287] Antibodies of the present invention can also be prepared by cellfree translation, as further described in Merk et al., J. Biochem.(Tokyo) 125(2): 328-33 (1999) and Ryabova et al., Nature Biotechnol.15(1): 79-84 (1997), and in the milk of transgenic animals, as furtherdescribed in Pollock et al., J. Inmunol. Methods 231(1-2): 147-57(1999), the disclosures of which are incorporated herein by reference intheir entireties.

[0288] The invention further provides antibody fragments that bindspecifically to one or more of the proteins and protein fragments of thepresent invention, to one or more of the proteins and protein fragmentsencoded by the isolated nucleic acids of the present invention, or thebinding of which can be competitively inhibited by one or more of theproteins and protein fragments of the present invention or one or moreof the proteins and protein fragments encoded by the isolated nucleicacids of the present invention.

[0289] Among such useful fragments are Fab, Fab′, Fv, F(ab)′₂, andsingle chain Fv (scFv) fragments. Other useful fragments are describedin Hudson, Curr. Opin. Biotechnol. 9(4): 395-402 (1998).

[0290] It is also an aspect of the present invention to provide antibodyderivatives that bind specifically to one or more of the proteins andprotein fragments of the present invention, to one or more of theproteins and protein fragments encoded by the isolated nucleic acids ofthe present invention, or the binding of which can be competitivelyinhibited by one or more of the proteins and protein fragments of thepresent invention or one or more of the proteins and protein fragmentsencoded by the isolated nucleic acids of the present invention.

[0291] Among such useful derivatives are chimeric, primatized, andhumanized antibodies; such derivatives are less immunogenic in humanbeings, and thus more suitable for in vivo administration, than areunmodified antibodies from non-human mammalian species. Another usefulderivative is PEGylation to increase the serum half life of theantibodies.

[0292] Chimeric antibodies typically include heavy and/or light chainvariable regions (including both CDR and framework residues) ofimmunoglobulins of one species, typically mouse, fused to constantregions of another species, typically human. See, e.g., U.S. Pat. No.5,807,715; Morrison et al., Proc. Natl. Acad. Sci USA.81(21): 6851-5(1984); Sharon et al., Nature 309(5966): 364-7 (1984); Takeda et al.,Nature 314(6010): 452-4 (1985), the disclosures of which areincorporated herein by reference in their entireties. Primatized andhumanized antibodies typically include heavy and/or light chain CDRsfrom a murine antibody grafted into a non-human primate or humanantibody V region framework, usually further comprising a human constantregion, Riechmann et al., Nature 332(6162): 323-7 (1988); Co et al.,Nature 351(6326): 501-2 (1991); U.S. Pat. Nos. 6,054,297; 5,821,337;5,770,196; 5,766,886; 5,821,123; 5,869,619; 6,180,377; 6,013,256;5,693,761; and 6,180,370, the disclosures of which are incorporatedherein by reference in their entireties.

[0293] Other useful antibody derivatives of the invention includeheteromeric antibody complexes and antibody fusions, such as diabodies(bispecific antibodies), single-chain diabodies, and intrabodies.

[0294] It is contemplated that the nucleic acids encoding the antibodiesof the present invention can be operably joined to other nucleic acidsforming a recombinant vector for cloning or for expression of theantibodies of the invention. The present invention includes anyrecombinant vector containing the coding sequences, or part thereof,whether for eukaryotic transduction, transfection or gene therapy. Suchvectors may be prepared using conventional molecular biology techniques,known to those with skill in the art, and would comprise DNA encodingsequences for the immunoglobulin V-regions including framework and CDRsor parts thereof, and a suitable promoter either with or without asignal sequence for intracellular transport. Such vectors may betransduced or transfected into eukaryotic cells or used for gene therapy(Marasco et al., Proc. Natl. Acad. Sci. (USA) 90: 7889-7893 (1993); Duanet al., Proc. Natl. Acad. Sci. (USA) 91: 5075-5079 (1994), byconventional techniques, known to those with skill in the art.

[0295] The antibodies of the present invention, including fragments andderivatives thereof, can usefully be labeled. It is, therefore, anotheraspect of the present invention to provide labeled antibodies that bindspecifically to one or more of the proteins and protein fragments of thepresent invention, to one or more of the proteins and protein fragmentsencoded by the isolated nucleic acids of the present invention, or thebinding of which can be competitively inhibited by one or more of theproteins and protein fragments of the present invention or one or moreof the proteins and protcin fragments encoded by the isolated nucleicacids of the present invention.

[0296] The choice of label depends, in part, upon the desired use.

[0297] For example, when the antibodies of the present invention areused for immunohistochemical staining of tissue samples, the label ispreferably an enzyme that catalyzes production and local deposition of adetectable product.

[0298] Enzymes typically conjugated to antibodies to permit theirimmunohistochemical visualization are well-known, and include alkalinephosphatase, β-galactosidase, glucose oxidase, horseradish peroxidase(HRP), and urease. Typical substrates for production and deposition ofvisually detectable products includeo-nitrophenyl-beta-D-galactopyranoside (ONPG); o-phenylenediaminedihydrochloride (OPD); p-nitrophenyl phosphate (PNPP);p-nitrophenyl-beta-D-galactopryanoside (PNPG); 3′,3′-diaminobenzidine(DAB); 3-amino-9-ethylcarbazole (AEC); 4-chloro-1-naphthol (CN);5-bromo-4-chloro-3-indolyl-phosphate (BCIP); ABTS®; BluoGal;iodonitrotetrazolium (INT); nitroblue tetrazolium chloride (NBT);phenazine methosulfate (PMS); phenolphlhalein monophosphate (PMP);tetramethyl benzidine (TMB); tetranitroblue tetrazolium (TNBT); X-Gal;X-Gluc; and X-Glucoside.

[0299] Other substrates can be used to produce products for localdeposition that are luminescent. For example, in the presence ofhydrogen peroxide (H₂O₂), horseradish peroxidase (HRP) can catalyze theoxidation of cyclic diacylhydrazides, such as luminol. Immediatelyfollowing the oxidation, the luminol is in an excited state(intermediate reaction product), which decays to the ground state byemitting light. Strong enhancement of the light emission is produced byenhancers, such as phenolic compounds. Advantages include highsensitivity, high resolution, and rapid detection without radioactivityand requiring only small amounts of antibody. See, e.g., Thorpe et al.,Methods Enzymol. 133: 331-53 (1986); Kricka et al., J. Immunoassay17(1): 67-83 (1996); and Lundqvist et al., J. Biolumin. Chemilumin.10(6): 353-9 (1995), the disclosures of which are incorporated herein byreference in their entireties. Kits for such enhanced chemiluminescentdetection (ECL) are available commercially.

[0300] The antibodies can also be labeled using colloidal gold.

[0301] As another example, when the antibodies of the present inventionare used, e.g., for flow cytometric detection, for scanning lasercytometric detection, or for fluorescent immunoassay, they can usefullybe labeled with fluorophores.

[0302] There are a wide variety of fluorophore labels that can usefullybe attached to the antibodies of the present invention.

[0303] For flow cytometric applications, both for extracellulardetection and for intracellular detection, common useful fluorophorescan be fluorescein isothiocyanate (FITC), allophycocyanin (APC),R-phycoerythrin (PE), peridinin chlorophyll protein (PerCP), Texas Red,Cy3, Cy5, fluorescence resonance energy tandem fluorophores such asPerCP-Cy5.5, PE-Cy5, PE-Cy5.5, PE-Cy7, PE-Texas Red, and APC-Cy7.

[0304] Other fluorophores include, inter alia, Alexa Fluor® 350, AlexaFluor® 488, Alexa Fluor® 532, Alexa Fluor® 546, Alexa Fluor® 568, AlexaFluor® 594, Alexa Fluor® 647 (monoclonal antibody labeling kitsavailable from Molecular Probes, Inc., Eugene, Oreg., USA), BODIPY dyes,such as BODIPY 493/503, BODIPY FL, BODIPY R6G, BODIPY 530/550, BODIPYTMR, BODIPY 558/568, BODIPY 558/568, BODIPY 564/570, BODIPY 576/589,BODIPY 581/591, BODIPY TR, BODIPY 630/650, BODIPY 650/665, Cascade Blue,Cascade Yellow, Dansyl, lissamine rhodaminc B, Marina Blue, Oregon Green488, Oregon Green 514, Pacific Blue, rhodamine 6G, rhodamine green,rhodamine red, tetramethylrhodamine, Texas Red (available from MolecularProbes, Inc., Eugene, Oreg., USA), and Cy2, Cy3, Cy3.5, Cy5, Cy5.5, Cy7,all of which are also useful for fluorescently labeling the antibodiesof the present invention.

[0305] For secondary detection using labeled avidin, streptavidin,captavidin or neutravidin, the antibodies of the present invention canusefully be labeled with biotin.

[0306] When the antibodies of the present invention are used, e.g., forWestern blotting applications, they can usefully be labeled withradioisotopes, such as ³³P, ³²P, ³⁵S, ³H, and ¹²⁵I.

[0307] As another example, when the antibodies of the present inventionare used for radioimmunothcrapy, the label can usefully be ²²⁸Th, ²²⁷Ac,²²⁵Ac, ²²³Ra, ²¹³Bi, ²¹²Pb, ²¹²Bi, 211 At, ²⁰³Pb, ¹⁹⁴Os, ¹⁸⁸Re, ¹⁸⁶Re,¹⁵³Sm, ¹⁴⁹Tb, ¹³¹I, ¹²⁵I, ¹¹¹In, ¹⁰⁵Rh, ^(99m)Tc, ⁹⁷Ru, ⁹⁰y, ⁹⁰Sr, ⁸⁸Y,⁷²Se, ⁶⁷Cu, or ⁴⁷Sc.

[0308] As another example, when the antibodies of the present inventionare to be used for in vivo diagnostic use, they can be rendereddetectable by conjugation to MRI contrast agents, such as gadoliniumdiethylenctriamincpcntaacctic acid (DTPA), Lauffer et al., Radiology207(2): 529-38 (1998), or by radioisotopic labeling.

[0309] As would be understood, use of the labels described above is notrestricted to the application for which they are mentioned.

[0310] The antibodies of the present invention, including fragments andderivatives thereof, can also be conjugated to toxins, in order totarget the toxin's ablative action to cells that display and/or expressthe proteins of the present invention. Commonly, the antibody in suchimmunotoxins is conjugated to Pseudomonas exotoxin A, diphtheria toxin,shiga toxin A, anthrax toxin lethal factor, or ricin. See Hall (ed.),Immunotoxin Methods and Protocols (Methods in Molecular Biology, vol.166), Humana Press (2000); and Frankel et al. (eds.), ClinicalApplications of Immunotoxins, Springer-Verlag (1998), the disclosures ofwhich are incorporated herein by reference in their entireties.

[0311] The antibodies of the present invention can usefully be attachedto a substrate, and it is, therefore, another aspect of the invention toprovide antibodies that bind specifically to one or more of the proteinsand protein fragments of the present invention, to one or more of theproteins and protein fragments encoded by the isolated nucleic acids ofthe present invention, or the binding of which can be competitivelyinhibited by one or more of the proteins and protein fragments of thepresent invention or one or more of the proteins and protein fragmentsencoded by the isolated nucleic acids of the present invention, attachedto a substrate.

[0312] Substrates can be porous or nonporous, planar or nonplanar.

[0313] For example, the antibodies of the present invention can usefullybe conjugated to filtration media, such as NHS-activated Sepharose orCNBr-activated Sepharose for purposes of immunoaffinity chromatography.

[0314] For example, the antibodies of the present invention can usefullybe attached to paramagnetic microspheres, typically bybiotin-streptavidin interaction, which microspheres can then be used forisolation of cells that express or display the proteins of the presentinvention. As another example, the antibodies of the present inventioncan usefully be attached to the surface of a microtiter plate for ELISA.

[0315] As noted above, the antibodies of the present invention can beproduced in prokaryotic and eukaryotic cells. It is, therefore, anotheraspect of the present invention to provide cells that express theantibodies of the present invention, including hybridoma cells, B cells,plasma cells, and host cells recombinantly modified to express theantibodies of the present invention.

[0316] In yet a further aspect, the present invention provides aptamersevolved to bind specifically to one or more of the proteins and proteinfragments of the present invention, to one or more of the proteins andprotein fragments encoded by the isolated nucleic acids of the presentinvention, or the binding of which can be competitively inhibited by oneor more of the proteins and protein fragments of the present inventionor one or more of the proteins and protein fragments encoded by theisolated nucleic acids of the present invention.

[0317] In sum, one of skill in the art, provided with the teachings ofthis invention, has available a variety of methods which may be used toalter the biological properties of the antibodies of this inventionincluding methods which would increase or decrease the stability orhalf-life, immunogenicity, toxicity, affinity or yield of a givenantibody molecule, or to alter it in any other way that may render itmore suitable for a particular application.

[0318] Transgenic Animals and Cells

[0319] In another aspect, the invention provides transgenic cells andnon-human organisms comprising nucleic acid molecules of the invention.In a preferred embodiment, the transgenic cells and non-human organismscomprise a nucleic acid molecule encoding a BSP. In a preferredembodiment, the BSP comprises an amino acid sequence selected from SEQID NO: 160 through 282, or a fragment, mutein, homologous protein orallelic variant thereof. In another preferred embodiment, the transgeniccells and non-human organism comprise a BSNA of the invention,preferably a BSNA comprising a nucleotide sequence selected from thegroup consisting of SEQ ID NO: 1 through 159, or a part, substantiallysimilar nucleic acid molecule, allelic variant or hybridizing nucleicacid molecule thereof.

[0320] In another embodiment, the transgenic cells and non-humanorganisms have a targeted disruption or replacement of the endogenousorthologue of the human BSG. The transgenic cells can be embryonic stemcells or somatic cells. The transgenic non-human organisms can bechimeric, nonchimeric heterozygotes, and nonchimeric homozygotes.Methods of producing transgenic animals are well-known in the art. See,e.g., Hogan et al., Manipulating the Mouse Embryo: A Laboratory Manual,2d ed., Cold Spring Harbor Press (1999); Jackson et al., Mouse Geneticsand Transgenics: A Practical Approach, Oxford University Press (2000);and Pinkert, Transgenic Animal Technology: A Laboratory Handbook,Academic Press (1999).

[0321] Any technique known in the art may be used to introduce a nucleicacid molecule of the invention into an animal to produce the founderlines of transgenic animals. Such techniques include, but are notlimited to, pronuclear microinjection. (see, e.g., Paterson et al.,Appl. Microbiol. Biotechnol. 40: 691-698 (1994); Carver et al.,Biotechnology 11: 1263-1270 (1993); Wright et al., Biotechnology 9:830-834 (1991); and U.S. Pat. No. 4,873,191 (1989 retrovirus-mediatedgene transfer into germ lines, blastocysts or embryos (see, e.g., Vander Putten et al., Proc. Natl. Acad. Sci., USA 82: 6148-6152 (1985));gene targeting in embryonic stem cells (see, e.g., Thompson et al., Cell56: 313-321 (1989)); electroporation of cells or embryos (see, e.g., Lo,1983, Mol. Cell. Biol. 3: 1803-1814 (1983)); introduction using a genegun (see, e.g., Ulmer et al., Science 259: 1745-49 (1993); introducingnucleic acid constructs into embryonic pleuripotent stem cells andtransferring the stem cells back into the blastocyst; and spcrm-mediatedgene transfer (see, e.g., Lavitrano et al., Cell 57: 717-723 (1989)).

[0322] Other techniques include, for example, nuclear transfer intoenucleated oocytes of nuclei from cultured embryonic, fetal, or adultcells induced to quiescence (see, e.g., Campell et al., Nature 380:64-66 (1996); Wilmut et al., Nature 385: 810-813 (1997)). The presentinvention provides for transgenic animals that carry the transgene(i.e., a nucleic acid molecule of the invention) in all their cells, aswell as animals which carry the transgene in some, but not all theircells, i. e., mosaic animals or chimeric animals.

[0323] The transgene may be integrated as a single transgene or asmultiple copies, such as in concatamers, e.g., head-to-head tandems orhead-to-tail tandems. The transgene may also be selectively introducedinto and activated in a particular cell type by following, e.g., theteaching of Lasko et al. et al., Proc. Natl. Acad. Sci. USA 89:6232-6236 (1992). The regulatory sequences required for such a cell-typespecific activation will depend upon the particular cell type ofinterest, and will be apparent to those of skill in the art.

[0324] Once transgenic animals have been generated, the expression ofthe recombinant gene may be assayed utilizing standard techniques.Initial screening may be accomplished by Southern blot analysis or PCRtechniques to analyze animal tissues to verify that integration of thetransgene has taken place. The level of mRNA expression of the transgenein the tissues of the transgenic animals may also be assessed usingtechniques which include, but are not limited to, Northern blot analysisof tissue samples obtained from the animal, in situ hybridizationanalysis, and reverse transcriptase-PCR (RT-PCR). Samples of transgenicgene-expressing tissue may also be evaluated immunocytochemically orimmunohistochemically using antibodies specific for the transgeneproduct.

[0325] Once the founder animals are produced, they may be bred, inbred,outbred, or crossbred to produce colonies of the particular animal.Examples of such breeding strategies include, but are not limited to:outbreeding of founder animals with more than one integration site inorder to establish separate lines; inbreeding of separate lines in orderto produce compound transgenics that express the transgene at higherlevels because of the effects of additive expression of each transgene;crossing of heterozygous transgenic animals to produce animalshomozygous for a given integration site in order to both augmentexpression and eliminate the need for screening of animals by DNAanalysis; crossing of separate homozygous lines to produce compoundheterozygous or homozygous lines; and breeding to place the transgene ona distinct background that is appropriate for an experimental model ofinterest.

[0326] Transgenic animals of the invention have uses which include, butare not limited to, animal model systems useful in elaborating thebiological function of polypeptides of the present invention, studyingconditions and/or disorders associated with aberrant expression, and inscreening for compounds effective in ameliorating such conditions and/ordisorders.

[0327] Methods for creating a transgenic animal with a disruption of atargeted gene are also well-known in the art. In general, a vector isdesigned to comprise some nucleotide sequences homologous to theendogenous targeted gene. The vector is introduced into a cell so thatit may integrate, via homologous recombination with chromosomalsequences, into the endogenous gene, thereby disrupting the function ofthe endogenous gene. The transgene may also be selectively introducedinto a particular cell type, thus inactivating the endogenous gene inonly that cell type. See, e.g., Gu et al., Science 265: 103-106 (1994).The regulatory sequences required for such a cell-type specificinactivation will depend upon the particular cell type of interest, andwill be apparent to those of skill in the art. See, e.g., Smithies etal., Nature 317: 230-234 (1985); Thomas et al., Cell 51: 503-512 (1987);Thompson et al., Cell 5: 313-321 (1989).

[0328] In one embodiment, a mutant, non-functional nucleic acid moleculeof the invention (or a completely unrelated DNA sequence) flanked by DNAhomologous to the endogenous nucleic acid sequence (either the codingregions or regulatory regions of the gene) can be used, with or withouta selectable marker and/or a negative selectable marker, to transfectcells that express polypeptides of the invention in vivo. In anotherembodiment, techniques known in the art are used to generate knockoutsin cells that contain, but do not express the gene of interest.Insertion of the DNA construct, via targeted homologous recombination,results in inactivation of the targeted gene. Such approaches areparticularly suited in research and agricultural fields wheremodifications to embryonic stem cells can be used to generate animaloffspring with an inactive targeted gene. See, e.g., Thomas, supra andThompson, supra. However this approach can be routinely adapted for usein humans provided the recombinant DNA constructs are directlyadministered or targeted to the required site in vivo using appropriateviral vectors that will be apparent to those of skill in the art.

[0329] In further embodiments of the invention, cells that aregenetically engineered to express the polypeptides of the invention, oralternatively, that are genetically engineered not to express thepolypeptides of the invention (e.g., knockouts) are administered to apatient in vivo. Such cells may be obtained from an animal or patient oran MHC compatible donor and can include, but are not limited tofibroblasts, bone marrow cells, blood cells (e.g., lymphocytes),adipocytes, muscle cells, endothelial cells etc. The cells aregenetically engineered in vitro using recombinant DNA techniques tointroduce the coding sequence of polypeptides of the invention into thecells, or alternatively, to disrupt the coding sequence and/orendogenous regulatory sequence associated with the polypeptides of theinvention, e.g., by transduction (using viral vectors, and preferablyvectors that integrate the transgene into the cell genome) ortransfection procedures, including, but not limited to, the use ofplasmids, cosmids, YACs, naked DNA, electroporation, liposomes, etc.

[0330] The coding sequence of the polypeptides of the invention can beplaced under the control of a strong constitutive or inducible promoteror promoter/enhancer to achieve expression, and preferably secretion, ofthe polypeptides of the invention. The engineered cells which expressand preferably secrete the polypeptides of the invention can beintroduced into the patient systemically, e.g., in the circulation, orintraperitoneally.

[0331] Alternatively, the cells can be incorporated into a matrix andimplanted in the body, e.g., genetically engineered fibroblasts can beimplanted as part of a skin graft; genetically engineered endothelialcells can be implanted as part of a lymphatic or vascular graft. See,e.g., U.S. Pat. Nos. 5,399,349 and 5,460,959, each of which isincorporated by reference herein in its entirety.

[0332] When the cells to be administered are non-autologous or non-MHCcompatible cells, they can be administered using well-known techniqueswhich prevent the development of a host immune response against theintroduced cells. For example, the cells may be introduced in anencapsulated form which, while allowing for an exchange of componentswith the immediate extracellular environment, does not allow theintroduced cells to be recognized by the host immune system.

[0333] Transgenic and “knock-out” animals of the invention have useswhich include, but are not limited to, animal model systems useful inelaborating the biological function of polypeptides of the presentinvention, studying conditions and/or disorders associated with aberrantexpression, and in screening for compounds effective in amelioratingsuch conditions and/or disorders.

[0334] Computer Readable Means

[0335] A further aspect of the invention relates to a computer readablemeans for storing the nucleic acid and amino acid sequences of theinstant invention. In a preferred embodiment, the invention provides acomputer readable means for storing SEQ ID NO: 1 through 159 and SEQ IDNO: 160 through 282 as described herein, as the complete set ofsequences or in any combination. The records of the computer readablemeans can be accessed for reading and display and for interface with acomputer system for the application of programs allowing for thelocation of data upon a query for data meeting certain criteria, thecomparison of sequences, the alignment or ordering of sequences meetinga set of criteria, and the like.

[0336] The nucleic acid and amino acid sequences of the invention areparticularly useful as components in databases useful for searchanalyses as well as in sequence analysis algorithms. As used herein, theterms “nucleic acid sequences of the invention” and “amino acidsequences of the invention” mean any detectable chemical or physicalcharacteristic of a polynucleotide or polypeptide of the invention thatis or may be reduced to or stored in a computer readable form. Theseinclude, without limitation, chromatographic scan data or peak data,photographic data or scan data therefrom, and mass spectrographic data.

[0337] This invention provides computer readable media having storedthereon sequences of the invention. A computer readable medium maycomprise one or more of the following: a nucleic acid sequencecomprising a sequence of a nucleic acid sequence of the invention; anamino acid sequence comprising an amino acid sequence of the invention;a set of nucleic acid sequences wherein at least one of said sequencescomprises the sequence of a nucleic acid sequence of the invention; aset of amino acid sequences wherein at least one of said sequencescomprises the sequence of an amino acid sequence of the invention; adata set representing a nucleic acid sequence comprising the sequence ofone or more nucleic acid sequences of the invention; a data setrepresenting a nucleic acid sequence encoding an amino acid sequencecomprising the sequence of an amino acid sequence of the invention; aset of nucleic acid sequences wherein at least one of said sequencescomprises the sequence of a nucleic acid sequence of the invention; aset of amino acid sequences wherein at least one of said sequencescomprises the sequence of an amino acid sequence of the invention; adata set representing a nucleic acid sequence comprising the sequence ofa nucleic acid sequence of the invention; a data set representing anucleic acid sequence encoding an amino acid sequence comprising thesequence of an amino acid sequence of the invention. The computerreadable medium can be any composition of matter used to storeinformation or data, including, for example, commercially availablefloppy disks, tapes, hard drives, compact disks, and video disks.

[0338] Also provided by the invention are methods for the analysis ofcharacter sequences, particularly genetic sequences. Preferred methodsof sequence analysis include, for example, methods of sequence homologyanalysis, such as identity and similarity analysis, RNA structureanalysis, sequence assembly, cladistic analysis, sequence motifanalysis, open reading frame determination, nucleic acid base calling,and sequencing chromatogram peak analysis.

[0339] A computer-based method is provided for performing nucleic acidsequence identity or similarity identification. This method comprisesthe steps of providing a nucleic acid sequence comprising the sequenceof a nucleic acid of the invention in a computer readable medium; andcomparing said nucleic acid sequence to at least one nucleic acid oramino acid sequence to identify sequence identity or similarity.

[0340] A computer-based method is also provided for performing aminoacid homology identification, said method comprising the steps of:providing an amino acid sequence comprising the sequence of an aminoacid of the invention in a computer readable medium; and comparing saidan amino acid sequence to at least one nucleic acid or an amino acidsequence to identify homology.

[0341] A computer-based method is still further provided for assembly ofoverlapping nucleic acid sequences into a single nucleic acid sequence,said method comprising the steps of: providing a first nucleic acidsequence comprising the sequence of a nucleic acid of the invention in acomputer readable medium; and screening for at least one overlappingregion between said first nucleic acid sequence and a second nucleicacid sequence.

[0342] Diagnostic Methods for Breast Cancer

[0343] The present invention also relates to quantitative andqualitative diagnostic assays and methods for detecting, diagnosing,monitoring, staging and predicting cancers by comparing expression of aBSNA or a BSP in a human patient that has or may have breast cancer, orwho is at risk of developing breast cancer, with the expression of aBSNA or a BSP in a normal human control. For purposes of the presentinvention, “expression of a BSNA” or “BSNA expression” means thequantity of BSG mRNA that can be measured by any method known in the artor the level of transcription that can be measured by any method knownin the art in a cell, tissue, organ or whole patient. Similarly, theterm “expression of a BSP” or “BSP expression” means the amount of BSPthat can be measured by any method known in the art or the level oftranslation of a BSG BSNA that can be measured by any method known inthe art.

[0344] The present invention provides methods for diagnosing breastcancer in a patient, in particular squamous cell carcinoma, by analyzingfor changes in levels of BSNA or BSP in cells, tissues, organs or bodilyfluids compared with levels of BSNA or BSP in cells, tissues, organs orbodily fluids of preferably the same type from a normal human control,wherein an increase, or decrease in certain cases, in levels of a BSNAor BSP in the patient versus the normal human control is associated withthe presence of breast cancer or with a predilection to the disease. Inanother preferred embodiment, the present invention provides methods fordiagnosing breast cancer in a patient by analyzing changes in thestructure of the mRNA of a BSG compared to the mRNA from a normalcontrol. These changes include, without limitation, aberrant splicing,alterations in polyadenylation and/or alterations in 5′ nucleotidecapping. In yet another preferred embodiment, the present inventionprovides methods for diagnosing breast cancer in a patient by analyzingchanges in a BSP compared to a BSP from a normal control. These changesinclude, e.g., alterations in glycosylation and/or phosphorylation ofthe BSP or subcellular BSP localization.

[0345] In a preferred embodiment, the expression of a BSNA is measuredby determining the amount of an mRNA that encodes an amino acid sequenceselected from SEQ ID NO: 160 through 282, a homolog, an allelic variant,or a fragment thereof. In a more preferred embodiment, the BSNAexpression that is measured is the level of expression of a BSNA mRNAselected from SEQ ID NO: 1 through 159, or a hybridizing nucleic acid,homologous nucleic acid or allelic variant thereof, or a part of any ofthese nucleic acids. BSNA expression may be measured by any method knownin the art, such as those described supra, including measuring mRNAexpression by Northern blot, quantitative or qualitative reversetranscriptase PCR (RT-PCR), microarray, dot or slot blots or in situhybridization. See, e.g., Ausubel (1992), supra; Ausubel (1999), supra;Sambrook (1989), supra; and Sambrook (2001), supra. BSNA transcriptionmay be measured by any method known in the art including using areporter gene hooked up to the promoter of a BSG of interest or doingnuclear run-off assays. Alterations in mRNA structure, e.g., aberrantsplicing variants, may be determined by any method known in the art,including, RT-PCR followed by sequencing or restriction analysis. Asnecessary, BSNA expression may be compared to a known control, such asnormal breast nucleic acid, to detect a change in expression.

[0346] In another preferred embodiment, the expression of a BSP ismeasured by determining the level of a BSP having an amino acid sequenceselected from the group consisting of SEQ ID NO: 160 through 282, ahomolog, an allclic variant, or a fragment thereof. Such levels arepreferably determined in at least one of cells, tissues, organs and/orbodily fluids, including determination of normal and abnormal levels.Thus, for instance, a diagnostic assay in accordance with the inventionfor diagnosing over- or underexpression of BSNA or BSP compared tonormal control bodily fluids, cells, or tissue samples may be used todiagnose the presence of breast cancer. The expression level of a BSPmay be determined by any method known in the art, such as thosedescribed supra. In a preferred embodiment, the BSP expression level maybe determined by radioimmunoassays, competitive-binding assays, ELISA,Western blot, FACS, immunohistochemistry, immunoprecipitation, proteomicapproaches: two-dimensional gel electrophoresis (2D electrophoresis) andnon-gel-based approaches such as mass spectrometry or proteininteraction profiling. See, e.g, Harlow (1999), supra; Ausubel (1992),supra; and Ausubel (1999), supra. Alterations in the BSP structure maybe determined by any method known in the art, including, e.g., usingantibodies that specifically recognize phosphoserine, phosphothreonineor phosphotyrosinc residues, two-dimensional polyacrylamide gelelectrophoresis (2D PAGE) and/or chemical analysis of amino acidresidues of the protein. Id.

[0347] In a preferred embodiment, a radioimmunoassay (RIA) or an ELISAis used. An antibody specific to a BSP is prepared if one is not alreadyavailable. In a preferred embodiment, the antibody is a monoclonalantibody. The anti-BSP antibody is bound to a solid support and any freeprotein binding sites on the solid support are blocked with a proteinsuch as bovine serum albumin. A sample of interest is incubated with theantibody on the solid support under conditions in which the BSP willbind to the anti-BSP antibody. The sample is removed, the solid supportis washed to remove unbound material, and an anti-BSP antibody that islinked to a detectable reagent (a radioactive substance for RIA and anenzyme for ELISA) is added to the solid support and incubated underconditions in which binding of the BSP to the labeled antibody willoccur. After binding, the unbound labeled antibody is removed bywashing. For an ELISA, one or more substrates are added to produce acolored reaction product that is based upon the amount of a BSP in thesample. For an RIA, the solid support is counted for radioactive decaysignals by any method known in the art. Quantitative results for bothRIA and ELISA typically are obtained by reference to a standard curve.

[0348] Other methods to measure BSP levels arc known in the art. Forinstance, a competition assay may be employed wherein an anti-BSPantibody is attached to a solid support and an allocated amount of alabeled BSP and a sample of interest are incubated with the solidsupport. The amount of labeled BSP detected which is attached to thesolid support can be correlated to the quantity of a BSP in the sample.

[0349] Of the proteomic approaches, 2D PAGE is a well-known technique.Isolation of individual proteins from a sample such as serum isaccomplished using sequential separation of proteins by isoelectricpoint and molecular weight. Typically, polypeptides are first separatedby isoelectric point (the first dimension) and then separated by sizeusing an electric current (the second dimension). In general, the seconddimension is perpendicular to the first dimension. Because no twoproteins with different sequences are identical on the basis of bothsize and charge, the result of 2D PAGE is a roughly square gel in whicheach protein occupies a unique spot. Analysis of the spots with chemicalor antibody probes, or subsequent protein microsequencing can reveal therelative abundance of a given protein and the identity of the proteinsin the sample.

[0350] Expression levels of a BSNA can bc determined by any method knownin the art, including PCR and other nucleic acid methods, such as ligasechain reaction (LCR) and nucleic acid sequence based amplification(NASBA), can be used to detect malignant cells for diagnosis andmonitoring of various malignancies. For example, reverse-transcriptasePCR (RT-PCR) is a powerful technique which can be used to detect thepresence of a specific mRNA population in a complex mixture of thousandsof other mRNA species. In RT-PCR, an mRNA species is first reversetranscribed to complementary DNA (cDNA) with use of the enzyme reversetranscriptase; the cDNA is then amplified as in a standard PCR reaction.

[0351] Hybridization to specific DNA molecules (e.g., oligonucleotides)arrayed on a solid support can be used to both detect the expression ofand quantitate the level of expression of one or more BSNAs of interest.In this approach, all or a portion of one or more BSNAs is fixed to asubstrate. A sample of interest, which may comprise RNA, e.g., total RNAor polyA-selected mRNA, or a complementary DNA (cDNA) copy of the RNA isincubated with the solid support under conditions in which hybridizationwill occur between the DNA on the solid support and the nucleic acidmolecules in the sample of interest. Hybridization between thesubstrate-bound DNA and the nucleic acid molecules in the sample can bedetected and quantitated by several means, including, withoutlimitation, radioactive labeling or fluorescent labeling of the nucleicacid molecule or a secondary molecule designed to detect the hybrid.

[0352] The above tests can be carried out on samples derived from avariety of cells, bodily fluids and/or tissue extracts such ashomogenates or solubilized tissue obtained from a patient. Tissueextracts are obtained routinely from tissue biopsy and autopsy material.Bodily fluids useful in the present invention include blood, urine,saliva or any other bodily secretion or derivative thereof. By blood itis meant to include whole blood, plasma, serum or any derivative ofblood. In a preferred embodiment, the specimen tested for expression ofBSNA or BSP includes, without limitation, breast tissue, fluid obtainedby bronchial alveolar lavage (BAL), sputum, breast cells grown in cellculture, blood, serum, lymph node tissue and lymphatic fluid. In anotherpreferred embodiment, especially when metastasis of a primary breastcancer is known or suspected, specimens include, without limitation,tissues from brain, bone, bone marrow, liver, adrenal glands and colon.In general, the tissues may be sampled by biopsy, including, withoutlimitation, needle biopsy, e.g., transthoracic ncedlc aspiration,ccrvical mediatinoscopy, endoscopic lymph node biopsy, video-assistedthoracoscopy, exploratory thoracotomy, bone marrow biopsy and bonemarrow aspiration. See Scott, supra and Franklin, pp. 529-570, in Kane,supra. For early and inexpensive detection, assaying for changes inBSNAs or BSPs in cells in sputum samples may be particularly useful.Methods of obtaining and analyzing sputum samples is disclosed inFranklin, supra.

[0353] All the methods of the present invention may optionally includedetermining the expression levels of one or more other cancer markers inaddition to determining the expression level of a BSNA or BSP. In manycases, the use of another cancer marker will decrease the likelihood offalse positives or false negatives. In one embodiment, the one or moreother cancer markers include other BSNA or BSPs as disclosed herein.Other cancer markers useful in the present invention will depend on thecancer being tested and are known to those of skill in the art. In apreferred embodiment, at least one other cancer marker in addition to aparticular BSNA or BSP is measured. In a more preferred embodiment, atleast two other additional cancer markers are used. In an even morepreferred embodiment, at least three, more preferably at least five,even more preferably at least ten additional cancer markers are used.

[0354] Diagnosing

[0355] In one aspect, the invention provides a method for determiningthe expression levels and/or structural alterations of one or more BSNAsand/or BSPs in a sample from a patient suspected of having breastcancer. In general, the method comprises the steps of obtaining thesample from the patient, determining the expression level or structuralalterations of a BSNA and/or BSP and then ascertaining whether thepatient has breast cancer from the expression level of the BSNA or BSP.In general, if high expression relative to a control of a BSNA or BSP isindicative of breast cancer, a diagnostic assay is considered positiveif the level of expression of the BSNA or BSP is at least two timeshigher, and more preferably are at least five times higher, even morepreferably at least ten times higher, than in preferably the same cells,tissues or bodily fluid of a normal human control. In contrast, if lowexpression relative to a control of a BSNA or BSP is indicative ofbreast cancer, a diagnostic assay is considered positive if the level ofexpression of the BSNA or BSP is at least two times lower, morepreferably are at least five times lower, even more preferably at leastten times lower than in preferably the same cells, tissues or bodilyfluid of a normal human control. The normal human control may be from adifferent patient or from uninvolved tissue of the same patient.

[0356] The present invention also provides a method of determiningwhether breast cancer has metastasized in a patient. One may identifywhether the breast cancer has metastasized by measuring the expressionlevels and/or structural alterations of one or more BSNAs and/or BSPs ina variety of tissues. The presence of a BSNA or BSP in a certain tissueat levels higher than that of corresponding noncancerous tissue (e.g.,the same tissue from another individual) is indicative of metastasis ifhigh level expression of a BSNA or BSP is associated with breast cancer.Similarly, the presence of a BSNA or BSP in a tissue at levels lowerthan that of corresponding noncancerous tissue is indicative ofmetastasis if low level expression of a BSNA or BSP is associated withbreast cancer. Further, the presence of a structurally altered BSNA orBSP that is associated with breast cancer is also indicative ofmetastasis.

[0357] In general, if high expression relative to a control of a BSNA orBSP is indicative of metastasis, an assay for metastasis is consideredpositive if the level of expression of the BSNA or BSP is at least twotimes higher, and more preferably are at least five times higher, evenmore preferably at least ten times higher, than in preferably the samecells, tissues or bodily fluid of a nonnal human control. In contrast,if low expression relative to a control of a BSNA or BSP is indicativeof metastasis, an assay for metastasis is considered positive if thelevel of expression of the BSNA or BSP is at least two times lower, morepreferably are at least five times lower, even more preferably at leastten times lower than in preferably the same cells, tissues or bodilyfluid of a normal human control.

[0358] The BSNA or BSP of this invention may be used as element in anarray or a multi-analyte test to recognize expression patternsassociated with breast cancers or other breast related disorders. Inaddition, the sequences of either the nucleic acids or proteins may beused as elements in a computer program for pattern recognition of breastdisorders.

[0359] Staging

[0360] The invention also provides a method of staging breast cancer ina human patient. The method comprises identifying a human patient havingbreast cancer and analyzing cells, tissues or bodily fluids from suchhuman patient for expression levels and/or structural alterations of oneor more BSNAs or BSPs. First, one or more tumors from a variety ofpatients are staged according to procedures well-known in the art, andthe expression level of one or more BSNAs or BSPs is determined for eachstage to obtain a standard expression level for each BSNA and BSP. Then,the BSNA or BSP expression levels are determined in a biological samplefrom a patient whose stage of cancer is not known. The BSNA or BSPexpression levels from the patient are then compared to the standardexpression level. By comparing the expression level of the BSNAs andBSPs from the patient to the standard expression levels, one maydetermine the stage of the tumor. The same procedure may be followedusing structural alterations of a BSNA or BSP to determine the stage ofa breast cancer.

[0361] Monitoring

[0362] Further provided is a method of monitoring breast cancer in ahuman patient. One may monitor a human patient to determine whetherthere has been metastasis and, if there has been, when metastasis beganto occur. One may also monitor a human patient to determine whether apreneoplastic lesion has become cancerous. One may also monitor a humanpatient to determine whether a therapy, e.g., chemotherapy, radiotherapyor surgery, has decreased or eliminated the breast cancer. The methodcomprises identifying a human patient that one wants to monitor forbreast cancer, periodically analyzing cells, tissues or bodily fluidsfrom such human patient for expression levels of one or more BSNAs orBSPs, and comparing the BSNA or BSP levels over time to those BSNA orBSP expression levels obtained previously. Patients may also bemonitored by measuring one or more structural alterations in a BSNA orBSP that are associated with breast cancer.

[0363] If increased expression of a BSNA or BSP is associated withmetastasis, treatment failure, or conversion of a preneoplastic lesionto a cancerous lesion, then detecting an increase in the expressionlevel of a BSNA or BSP indicates that the tumor is metastasizing, thattreatment has failed or that the lesion is cancerous, respectively. Onehaving ordinary skill in the art would recognize that if this were thecase, then a decreased expression level would be indicative of nometastasis, effective therapy or failure to progress to a neoplasticlesion. If decreased expression of a BSNA or BSP is associated withmetastasis, treatment failure, or conversion of a preneoplastic lesionto a cancerous lesion, then detecting an decrease in the expressionlevel of a BSNA or BSP indicates that the tumor is metastasizing, thattreatment has failed or that the lesion is cancerous, respectively. In apreferred embodiment, the levels of BSNAs or BSPs are determined fromthe same cell type, tissue or bodily fluid as prior patient samples.Monitoring a patient for onset of breast cancer metastasis is periodicand preferably is done on a quarterly basis, but may be done more orless frequently.

[0364] The methods described herein can further be utilized asprognostic assays to identify subjects having or at risk of developing adisease or disorder associated with increased or decreased expressionlevels of a BSNA and/or BSP. The present invention provides a method inwhich a test sample is obtained from a human patient and one or moreBSNAs and/or BSPs are detected. The presence of higher (or lower) BSNAor BSP levels as compared to normal human controls is diagnostic for thehuman patient being at risk for developing cancer, particularly breastcancer. The effectiveness of therapeutic agents to decrease (orincrease) expression or activity of one or more BSNAs and/or BSPs of theinvention can also be monitored by analyzing levels of expression of theBSNAs and/or BSPs in a human patient in clinical trials or in in vitroscreening assays such as in human cells. In this way, the geneexpression pattern can serve as a marker, indicative of thephysiological response of the human patient or cells, as the case maybe, to the agent being tested.

[0365] Detection of Genetic Lesions or Mutations

[0366] The methods of the present invention can also be used to detectgenetic lesions or mutations in a BSG, thereby determining if a humanwith the genetic lesion is susceptible to developing breast cancer or todetermine what genetic lesions are responsible, or are partlyresponsible, for a person's existing breast cancer. Genetic lesions canbe detected, for example, by ascertaining the existence of a deletion,insertion and/or substitution of one or more nucleotides from the BSGsof this invention, a chromosomal rearrangement of BSG, an aberrantmodification of BSG (such as of the methylation pattern of the genomicDNA), or allelic loss of a BSG. Methods to detect such lesions in theBSG of this invention are known to those having ordinary skill in theart following the teachings of the specification.

[0367] Methods of Detecting Noncancerous Breast Diseases

[0368] The invention also provides a method for determining theexpression levels and/or structural alterations of one or more BSNAsand/or BSPs in a sample from a patient suspected of having or known tohave a noncancerous breast disease. In general, the method comprises thesteps of obtaining a sample from the patient, determining the expressionlevel or structural alterations of a BSNA and/or BSP, comparing theexpression level or structural alteration of the BSNA or BSP to a normalbreast control, and then ascertaining whether the patient has anoncancerous breast disease. In general, if high expression relative toa control of a BSNA or BSP is indicative of a particular noncancerousbreast disease, a diagnostic assay is considered positive if the levelof expression of the BSNA or BSP is at least two times higher, and morepreferably are at least five times higher, even more preferably at leastten times higher, than in preferably the same cells, tissues or bodilyfluid of a normal human control. In contrast, if low expression relativeto a control of a BSNA or BSP is indicative of a noncancerous breastdisease, a diagnostic assay is considered positive if the level ofexpression of the BSNA or BSP is at least two times lower, morepreferably are at least five times lower, even more preferably at leastten times lower than in preferably the same cells, tissues or bodilyfluid of a normal human control. The normal human control may be from adifferent patient or from uninvolved tissue of the same patient.

[0369] One having ordinary skill in the art may determine whether a BSNAand/or BSP is associated with a particular noncancerous breast diseaseby obtaining breast tissue from a patient having a noncancerous breastdisease of interest and determining which BSNAs and/or BSPs areexpressed in the tissue at either a higher or a lower level than innormal breast tissue. In another embodiment, one may determine whether aBSNA or BSP exhibits structural alterations in a particular noncancerousbreast disease state by obtaining breast tissue from a patient having anoncancerous breast disease of interest and determining the structuralalterations in one or more BSNAs and/or BSPs relative to normal breasttissue.

[0370] Methods for Identifying Breast Tissue

[0371] In another aspect, the invention provides methods for identifyingbreast tissue. These methods are particularly useful in, e.g., forensicscience, breast cell differentiation and development, and in tissueengineering.

[0372] In one embodiment, the invention provides a method fordetermining whether a sample is breast tissue or has breast tissue-likecharacteristics. The method comprises the steps of providing a samplesuspected of comprising breast tissue or having breast tissue-likecharacteristics, determining whether the sample expresses one or moreBSNAs and/or BSPs, and, if the sample expresses one or more BSNAs and/orBSPs, concluding that the sample comprises breast tissue. In a preferredembodiment, the BSNA encodes a polypeptide having an amino acid sequenceselected from SEQ ID NO: 160 through 282, or a homolog, allelic variantor fragment thereof. In a more preferred embodiment, the BSNA has anucleotide sequence selected from SEQ ID NO: 1 through 159, or ahybridizing nucleic acid, an allelic variant or a part thereof.Determining whether a sample expresses a BSNA can be accomplished by anymethod known in the art. Preferred methods include hybridization tomicroarrays, Northern blot hybridization, and quantitative orqualitative RT-PCR. In another preferred embodiment, the method can bepracticed by determining whether a BSP is expressed. Determining whethera sample expresses a BSP can be accomplished by any method known in theart. Preferred methods include Western blot, ELISA, RIA and 2D PAGE. Inone embodiment, the BSP has an amino acid sequence selected from SEQ IDNO: 160 through 282, or a homolog, allelic variant or fragment thereof.In another preferred embodiment, the expression of at least two BSNAsand/or BSPs is determined. In a more preferred embodiment, theexpression of at least three, more preferably four and even morepreferably five BSNAs and/or BSPs are determined.

[0373] In one embodiment, the method can be used to determine whether anunknown tissue is breast tissue. This is particularly useful in forensicscience, in which small, damaged pieces of tissues that are notidentifiable by microscopic or other means are recovered from a crime oraccident scene. In another embodiment, the method can be used todetermine whether a tissue is differentiating or developing into breasttissue. This is important in monitoring the effects of the addition ofvarious agents to cell or tissue culture, e.g., in producing new breasttissue by tissue engineering. These agents include, e.g., growth anddifferentiation factors, extracellular matrix proteins and culturemedium. Other factors that may be measured for effects on tissuedevelopment and differentiation include gene transfer into the cells ortissues, alterations in pH, aqueous:air interface and various otherculture conditions.

[0374] Methods for Producing and Modifying Breast Tissue

[0375] In another aspect, the invention provides methods for producingengineered breast tissue or cells. In one embodiment, the methodcomprises the steps of providing cells, introducing a BSNA or a BSG intothe cells, and growing the cells under conditions in which they exhibitone or more properties of breast tissue cells. In a preferredembodiment, the cells are pluripotent. As is well-known in the art,normal breast tissue comprises a large number of different cell types.Thus, in one embodiment, the engineered breast tissue or cells comprisesone of these cell types. In another embodiment, the engineered breasttissue or cells comprises more than one breast cell type. Further, theculture conditions of the cells or tissue may require manipulation inorder to achieve full differentiation and development of the breast celltissue. Methods for manipulating culture conditions are well-known inthe art.

[0376] Nucleic acid molecules encoding one or more BSPs are introducedinto cells, preferably pluripotent cells. In a preferred embodiment, thenucleic acid molecules encode BSPs having amino acid sequences selectedfrom SEQ ID NO: 160 through 282, or homologous proteins, analogs,allelic variants or fragments thereof. In a more preferred embodiment,the nucleic acid molecules have a nucleotide sequence selected from SEQID NO: 1 through 159, or hybridizing nucleic acids, allelic variants orparts thereof. In another highly preferred embodiment, a BSG isintroduced into the cells. Expression vectors and methods of introducingnucleic acid molecules into cells are well-known in the art and aredescribed in detail, supra.

[0377] Artificial breast tissue may be used to treat patients who havelost some or all of their breast function.

[0378] Pharmaceutical Compositions

[0379] In another aspect, the invention provides pharmaceuticalcompositions comprising the nucleic acid molecules, polypeptides,antibodies, antibody derivatives, antibody fragments, agonists,antagonists, and inhibitors of the present invention. In a preferredembodiment, the pharmaceutical composition comprises a BSNA or partthereof. In a more preferred embodiment, the BSNA has a nucleotidesequence selected from the group consisting of SEQ ID NO: 1 through 159,a nucleic acid that hybridizes thereto, an allelic variant thereof, or anucleic acid that has substantial sequence identity thereto. In anotherpreferred embodiment, the pharmaceutical composition comprises a BSP orfragment thereof. In a more preferred embodiment, the BSP having anamino acid sequence that is selected from the group consisting of SEQ IDNO: 160 through 282, a polypeptide that is homologous thereto, a fusionprotein comprising all or a portion of the polypeptide, or an analog orderivative thereof. In another preferred embodiment, the pharmaceuticalcomposition comprises an anti-BSP antibody, preferably an antibody thatspecifically binds to a BSP having an amino acid that is selected fromthe group consisting of SEQ ID NO: 160 through 282, or an antibody thatbinds to a polypeptide that is homologous thereto, a fusion proteincomprising all or a portion of the polypeptide, or an analog orderivative thereof.

[0380] Such a composition typically contains from about 0.1 to 90% byweight of a therapeutic agent of the invention formulated in and/or witha pharmaceutically acceptable carrier or excipient.

[0381] Pharmaceutical formulation is a well-established art, and isfurther described in Gennaro (ed.), Remington: The Science and Practiceof Pharmacy, 20^(th) ed., Lippincott, Williams & Wilkins (2000); Anselet al., Pharmaceutical Dosage Forms and Drug Delivery Systems, 7^(th)ed., Lippincott Williams & Wilkins (1999); and Kibbe (ed.), Handbook ofPharmaceutical Excipients American Pharmaceutical Association, 3^(rd)ed. (2000), the disclosures of which are incorporated herein byreference in their entireties, and thus need not be described in detailherein.

[0382] Briefly, formulation of the pharmaceutical compositions of thepresent invention will depend upon the route chosen for administration.The pharmaceutical compositions utilized in this invention can beadministered by various routes including both enteral and parenteralroutes, including oral, intravenous, intramuscular, subcutaneous,inhalation, topical, sublingual, rectal, intra-arterial, intramedullary,intrathecal, intraventricular, transmucosal, transdermal, intranasal,intraperitoneal, intrapulmonary, and intrauterine.

[0383] Oral dosage forms can be formulated as tablets, pills, dragees,capsules, liquids, gels, syrups, slurries, suspensions, and the like,for ingestion by the patient.

[0384] Solid formulations of the compositions for oral administrationcan contain suitable carriers or excipients, such as carbohydrate orprotein fillers, such as sugars, including lactose, sucrose, mannitol,or sorbitol; starch from corn, wheat, rice, potato, or other plants;cellulose, such as methyl cellulose, hydroxypropylmethyl-cellulose,sodium carboxymethylcellulose, or microcrystalline cellulose; gumsincluding arabic and tragacanth; proteins such as gelatin and collagen;inorganics, such as kaolin, calcium carbonate, dicalcium phosphate,sodium chloride; and other agents such as acacia and alginic acid.

[0385] Agents that facilitate disintegration and/or solubilization canbe added, such as the cross-linked polyvinyl pyrrolidone, agar, alginicacid, or a salt thereof, such as sodium alginate, microcrystallinecellulose, corn starch, sodium starch glycolate, and alginic acid.

[0386] Tablet binders that can be used include acacia, methylcellulose,sodium carboxymethylcellulose, polyvinylpyrrolidone (Povidone™),hydroxypropyl methylcellulose, sucrose, starch and ethylcellulose.

[0387] Lubricants that can be used include magnesium stearates, stearicacid, silicone fluid, talc, waxes, oils, and colloidal silica.

[0388] Fillers, agents that facilitate disintegration and/orsolubilization, tablet binders and lubricants, including theaforementioned, can be used singly or in combination.

[0389] Solid oral dosage forms need not be uniform throughout. Forexample, dragee cores can be used in conjunction with suitable coatings,such as concentrated sugar solutions, which can also contain gum arabic,talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/ortitanium dioxide, lacquer solutions, and suitable organic solvents orsolvent mixtures.

[0390] Oral dosage forms of the present invention include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a coating, such as glycerol or sorbitol. Push-fit capsulescan contain active ingredients mixed with a filler or binders, such aslactose or starches, lubricants, such as talc or magnesium stearate,and, optionally, stabilizers. In soft capsules, the active compounds canbe dissolved or suspended in suitable liquids, such as fatty oils,liquid, or liquid polyethylene glycol with or without stabilizers.

[0391] Additionally, dyestuffs or pigments can be added to the tabletsor dragee coatings for product identification or to characterize thequantity of active compound, i.e., dosage.

[0392] Liquid formulations of the pharmaceutical compositions for oral(enteral) administration arc prepared in water or other aqueous vehiclesand can contain various suspending agents such as methylcellulose,alginates, tragacanth, pectin, kelgin, carrageenan, acacia,polyvinylpyrrolidone, and polyvinyl alcohol. The liquid formulations canalso include solutions, emulsions, syrups and elixirs containing,together with the active compound(s), wetting agents, sweeteners, andcoloring and flavoring agents.

[0393] The pharmaceutical compositions of the present invention can alsobe formulated for parenteral administration. Formulations for parenteraladministration can be in the form of aqueous or non-aqueous isotonicsterile injection solutions or suspensions.

[0394] For intravenous injection, water soluble versions of thecompounds of the present invention are formulated in, or if provided asa lyophilate, mixed with, a physiologically acceptable fluid vehicle,such as 5% dextrose (“D5”), physiologically buffered saline, 0.9%saline, Hanks' solution, or Ringer's solution. Intravenous formulationsmay include carriers, excipients or stabilizers including, withoutlimitation, calcium, human senim albumin, citrate, acetate, calciumchloride, carbonate, and other salts.

[0395] Intramuscular preparations, e.g. a sterile formulation of asuitable soluble salt form of the compounds of the present invention,can be dissolved and administered in a pharmaceutical excipient such asWater-for-Injection, 0.9% saline, or 5% glucose solution. Alternatively,a suitable insoluble form of the compound can be prepared andadministered as a suspension in an aqueous base or a pharmaceuticallyacceptable oil base, such as an ester of a long chain fatty acid (e.g.,ethyl oleate), fatty oils such as sesame oil, triglycerides, orliposomes.

[0396] Parenteral formulations of the compositions can contain variouscarriers such as vegetable oils, dimethylacetamide, dimethylformamide,ethyl lactate, ethyl carbonate, isopropyl myristate, ethanol, polyols(glycerol, propylene glycol, liquid polyethylene glycol, and the like).

[0397] Aqueous injection suspensions can also contain substances thatincrease the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Non-lipid polycationic amino polymerscan also be used for delivery. Optionally, the suspension can alsocontain suitable stabilizers or agents that increase the solubility ofthe compounds to allow for the preparation of highly concentratedsolutions.

[0398] Pharmaceutical compositions of the present invention can also beformulated to permit injectable, long-term, deposition. Injectable depotforms may be made by forming microencapsulated matrices of the compoundin biodegradable polymers such as polylactide-polyglycolide. Dependingupon the ratio of drug to polymer and the nature of the particularpolymer employed, the rate of drug release can be controlled. Examplesof other biodegradable polymers include poly(orthoesters) andpoly(anhydrides). Depot injectable formulations are also prepared byentrapping the drug in microemulsions that are compatible with bodytissues.

[0399] The pharmaceutical compositions of the present invention can beadministered topically.

[0400] For topical use the compounds of the present invention can alsobe prepared in suitable forms to be applied to the skin, or mucusmembranes of the nose and throat, and can take the form of lotions,creams, ointments, liquid sprays or inhalants, drops, tinctures,lozenges, or throat paints. Such topical formulations further caninclude chemical compounds such as dimethylsulfoxide (DMSO) tofacilitate surface penetration of the active ingredient. In othertransdermal formulations, typically in patch-delivered formulations, thepharmaceutically active compound is formulated with one or more skinpenetrants, such as 2-N-methyl-pyrrolidone (NMP) or Azonc. A topicalsemi-solid ointment formulation typically contains a concentration ofthe active ingredient from about 1 to 20%, e.g., 5 to 10%, in a carriersuch as a pharmaceutical cream base.

[0401] For application to the eyes or ears, the compounds of the presentinvention can be presented in liquid or semi-liquid form formulated inhydrophobic or hydrophilic bases as ointments, creams, lotions, paintsor powders.

[0402] For rectal administration the compounds of the present inventioncan be administered in the form of suppositories admixed withconventional carriers such as cocoa butter, wax or other glyceride.

[0403] Inhalation formulations can also readily be formulated. Forinhalation, various powder and liquid formulations can be prepared. Foraerosol preparations, a sterile formulation of the compound or salt formof the compound may be used in inhalers, such as metered dose inhalers,and nebulizers. Aerosolized forms may be especially useful for treatingrespiratory disorders.

[0404] Alternatively, the compounds of the present invention can be inpowder form for reconstitution in the appropriate pharmaceuticallyacceptable carrier at the time of delivery.

[0405] The pharmaceutically active compound in the pharmaceuticalcompositions of the present invention can be provided as the salt of avariety of acids, including but not limited to hydrochloric, sulfuric,acetic, lactic, tartaric, malic, and succinic acid. Salts tend to bemore soluble in aqueous or other protonic solvents than are thecorresponding free base forms.

[0406] After pharmaceutical compositions have been prepared, they arepackaged in an appropriate container and labeled for treatment of anindicated condition.

[0407] The active compound will be present in an amount effective toachieve the intended purpose. The determination of an effective dose iswell within the capability of those skilled in the art.

[0408] A “therapeutically effective dose” refers to that amount ofactive ingredient, for example BSP polypeptide, fusion protein, orfragments thereof, antibodies specific for BSP, agonists, antagonists orinhibitors of BSP, which ameliorates the signs or symptoms of thedisease or prevents progression thereof; as would be understood in themedical arts, cure, although desired, is not required.

[0409] The therapeutically effective dose of the pharmaceutical agentsof the present invention can be estimated initially by in vitro tests,such as cell culture assays, followed by assay in model animals, usuallymice, rats, rabbits, dogs, or pigs. The animal model can also be used todetermine an initial preferred concentration range and route ofadministration.

[0410] For example, the ED50 (the dose therapeutically effective in 50%of the population) and LD50 (the dose lethal to 50% of the population)can be determined in one or more cell culture of animal model systems.The dose ratio of toxic to therapeutic effects is the therapeutic index,which can be expressed as LD50/ED50. Pharmaceutical compositions thatexhibit large therapeutic indices are preferred.

[0411] The data obtained from cell culture assays and animal studies areused in formulating an initial dosage range for human use, andpreferably provide a range of circulating concentrations that includesthe ED50 with little or no toxicity. After administration, or betweensuccessive administrations, the circulating concentration of activeagent varies within this range depending upon pharmacokinetic factorswell-known in the art, such as the dosage form employed, sensitivity ofthe patient, and the route of administration.

[0412] The exact dosage will be determined by the practitioner, in lightof factors specific to the subject requiring treatment. Factors that canbe taken into account by the practitioner include the severity of thedisease state, general health of the subject, age, weight, gender of thesubject, diet, time and frequency of administration, drugcombination(s), reaction sensitivities, and tolerance/response totherapy. Long-acting pharmaceutical compositions can be administeredevery 3 to 4 days, every week, or once every two weeks depending onhalf-life and clearance rate of the particular formulation.

[0413] Normal dosage amounts may vary from 0.1 to 100,000 micrograms, upto a total dose of about 1 g, depending upon the route ofadministration. Where the therapeutic agent is a protein or antibody ofthe present invention, the therapeutic protein or antibody agenttypically is administered at a daily dosage of 0.01 mg to 30 mg/kg ofbody weight of the patient (e.g., 1 mg/kg to 5 mg/kg). Thepharmaceutical formulation can be administered in multiple doses perday, if desired, to achieve the total desired daily dose.

[0414] Guidance as to particular dosages and methods of delivery isprovided in the literature and generally available to practitioners inthe art. Those skilled in the art will employ different formulations fornucleotides than for proteins or their inhibitors. Similarly, deliveryof polynucleotides or polypeptides will be specific to particular cells,conditions, locations, etc.

[0415] Conventional methods, known to those of ordinary skill in the artof medicine, can be used to administer the pharmaceutical formulation(s)of the present invention to the patient. The pharmaceutical compositionsof the present invention can be administered alone, or in combinationwith other therapeutic agents or interventions.

[0416] Therapeutic Methods

[0417] The present invention further provides methods of treatingsubjects having defects in a gene of the invention , e.g., inexpression, activity, distribution, localization, and/or solubility,which can manifest as a disorder of breast function. As used herein,“treating” includes all medically-acceptable types of therapeuticintervention, including palliation and prophylaxis (prevention) ofdisease. The term “treating” encompasses any improvement of a disease,including minor improvements. These methods are discussed below.

[0418] Gene Therapy and Vaccines

[0419] The isolated nucleic acids of the present invention can also beused to drive in vivo expression of the polypeptides of the presentinvention. In vivo expression can be driven from a vector, typically aviral vector, often a vector based upon a replication incompetentretrovirus, an adenovirus, or an adeno-associated virus (AAV), forpurpose of gene therapy. In vivo expression can also be driven fromsignals endogenous to the nucleic acid or from a vector, often a plasmidvector, such as pVAX1 (Invitrogen, Carlsbad, Calif., USA), for purposeof “naked” nucleic acid vaccination, as further described in U.S. Pat.Nos. 5,589,466; 5,679,647; 5,804,566; 5,830,877; 5,843,913; 5,880,104;5,958,891; 5,985,847; 6,017,897; 6,110,898; and 6,204,250, thedisclosures of which are incorporated herein by reference in theirentireties. For cancer therapy, it is preferred that the vector also betumor-selective. See, e.g., Doronin et al., J. Virol. 75: 3314-24(2001).

[0420] In another embodiment of the therapeutic methods of the presentinvention, a therapeutically effective amount of a pharmaceuticalcomposition comprising a nucleic acid of the present invention isadministered. The nucleic acid can be delivered in a vector that drivesexpression of a BSP, fusion protein, or fragment thereof, or withoutsuch vector. Nucleic acid compositions that can drive expression of aBSP are administered, for example, to complement a deficiency in thenative BSP, or as DNA vaccines. Expression vectors derived from virus,replication deficient retroviruses, adenovirus, adeno-associated (AAV)virus, herpes virus, or vaccinia virus can be used as can plasmids. See,e.g., Cid-Arregui, supra. In a preferred embodiment, the nucleic acidmolecule encodes a BSP having the amino acid sequence of SEQ ID NO: 160through 282, or a fragment, fusion protein, allelic variant or homologthereof.

[0421] In still other therapeutic methods of the present invention,pharmaceutical compositions comprising host cells that express a BSP,fusions, or fragments thereof can be administered. In such cases, thecells are typically autologous, so as to circumvent xenogeneic orallotypic rejection, and are administered to complement defects in BSPproduction or activity. In a preferred embodiment, the nucleic acidmolecules in the cells encode a BSP having the amino acid sequence ofSEQ ID NO: 160 through 282, or a fragment, fusion protein, allelicvariant or homolog thereof.

[0422] Antisense Administration

[0423] Antisense nucleic acid compositions, or vectors that driveexpression of a BSG antisense nucleic acid, are administered todownregulate transcription and/or translation of a BSG in circumstancesin which excessive production, or production of aberrant protein, is thepathophysiologic basis of disease.

[0424] Antisense compositions useful in therapy can have a sequence thatis complementary to coding or to noncoding regions of a BSG. Forexample, oligonucleotides derived from the transcription initiationsite, e.g., between positions −10 and +10 from the start site, arepreferred.

[0425] Catalytic antisense compositions, such as ribozymes, that arecapable of sequence-specific hybridization to BSG transcripts, are alsouseful in therapy. See, e.g., Phylactou, Adv. Drug Deliv. Rev. 44(2-3):97-108 (2000); Phylactou et al., Hum. Mol. Genet. 7(10): 1649-53 (1998);Rossi, Ciba Found. Symp. 209: 195-204 (1997); and Sigurdsson et al.,Trends Biotechnol. 13(8): 286-9 (1995), the disclosures of which areincorporated herein by reference in their entireties.

[0426] Other nucleic acids useful in the therapeutic methods of thepresent invention are those that are capable of triplex helix formationin or near the BSG gcnomic locus. Such triplexing oligonucleotides areable to inhibit transcription. See, e.g., Intody et al., Nucleic AcidsRes. 28(21): 4283-90 (2000); McGuffie et al., Cancer Res. 60(14): 3790-9(2000), the disclosures of which are incorporated herein by reference.Pharmaceutical compositions comprising such triplex forming oligos(TFOs) are administered in circumstances in which excessive production,or production of aberrant protein, is a pathophysiologic basis ofdisease.

[0427] In a preferred embodiment, the antisense molecule is derived froma nucleic acid molecule encoding a BSP, preferably a BSP comprising anamino acid sequence of SEQ ID NO: 160 through 282, or a fragment,allelic variant or homolog thereof. In a more preferred embodiment, theantisense molecule is derived from a nucleic acid molecule having anucleotide sequence of SEQ ID NO: 1 through 159, or a part, allelicvariant, substantially similar or hybridizing nucleic acid thereof.

[0428] Polypeptide Administration

[0429] In one embodiment of the therapeutic methods of the presentinvention, a therapeutically effective amount of a pharmaceuticalcomposition comprising a BSP, a fusion protein, fragment, analog orderivative thereof is administered to a subject with aclinically-significant BSP defect.

[0430] Protein compositions are administered, for example, to complementa deficiency in native BSP. In other embodiments, protein compositionsare administered as a vaccine to elicit a humoral and/or cellular immuneresponse to BSP. The immune response can be used to modulate activity ofBSP or, depending on the immunogen, to immunize against aberrant oraberrantly expressed forms, such as mutant or inappropriately expressedisoforms. In yet other embodiments, protein fusions having a toxicmoiety are administered to ablate cells that aberrantly accumulate BSP.

[0431] In a preferred embodiment, the polypeptide is a BSP comprising anamino acid sequence of SEQ ID NO: 160 through 282, or a fusion protein,allelic variant, homolog, analog or derivative thereof. In a morepreferred embodiment, the polypeptide is encoded by a nucleic acidmolecule having a nucleotide sequence of SEQ ID NO: 1 through 159, or apart, allelic variant, substantially similar or hybridizing nucleic acidthereof.

[0432] Antibody, Agonist and Antagonist Administration

[0433] In another embodiment of the therapeutic methods of the presentinvention, a therapeutically effective amount of a pharmaceuticalcomposition comprising an antibody (including fragment or derivativethereof) of the present invention is administered. As is well-known,antibody compositions are administered, for example, to antagonizeactivity of BSP, or to target therapeutic agents to sites of BSPpresence and/or accumulation. In a preferred embodiment, the antibodyspecifically binds to a BSP comprising an amino acid sequence of SEQ IDNO: 160 through 282, or a fusion protein, allelic variant, homolog,analog or derivative thereof. In a more preferred embodiment, theantibody specifically binds to a BSP encoded by a nucleic acid moleculehaving a nucleotide sequence of SEQ ID NO: 1 through 159, or a part,allelic variant, substantially similar or hybridizing nucleic acidthereof.

[0434] The present invention also provides methods for identifyingmodulators which bind to a BSP or have a modulatory effect on theexpression or activity of a BSP. Modulators which decrease theexpression or activity of BSP (antagonists) are believed to be useful intreating breast cancer. Such screening assays are known to those ofskill in the art and include, without limitation, cell-based assays andcell-free assays. Small molecules predicted via computer imaging tospecifically bind to regions of a BSP can also be designed, synthesizedand tested for use in the imaging and treatment of breast cancer.Further, libraries of molecules can be screened for potential anticanceragents by assessing the ability of the molecule to bind to the BSPsidentified herein. Molecules identified in the library as being capableof binding to a BSP are key candidates for further evaluation for use inthe treatment of breast cancer. In a preferred embodiment, thesemolecules will downregulate expression and/or activity of a BSP incells.

[0435] In another embodiment of the therapeutic methods of the presentinvention, a pharmaceutical composition comprising a non-antibodyantagonist of BSP is administered. Antagonists of BSP can be producedusing methods generally known in the art. In particular, purified BSPcan be used to screen libraries of pharmaceutical agents, oftencombinatorial libraries of small molecules, to identify those thatspecifically bind and antagonize at least one activity of a BSP.

[0436] In other embodiments a pharmaceutical composition comprising anagonist of a BSP is administered. Agonists can be identified usingmethods analogous to those used to identify antagonists.

[0437] In a preferred embodiment, the antagonist or agonist specificallybinds to and antagonizes or agonizes, respectively, a BSP comprising anamino acid sequence of SEQ ID NO: 160 through 282, or a fusion protein,allelic variant, homolog, analog or derivative thereof. In a morepreferred embodiment, the antagonist or agonist specifically binds toand antagonizes or agonizes, respectively, a BSP encoded by a nucleicacid molecule having a nucleotide sequence of SEQ ID NO: 1 through 159,or a part, allelic variant, substantially similar or hybridizing nucleicacid thereof.

[0438] Targeting Breast Tissue

[0439] The invention also provides a method in which a polypeptide ofthe invention, or an antibody thereto, is linked to a therapeutic agentsuch that it can be delivered to the breast or to specific cells in thebreast. In a preferred embodiment, an anti-BSP antibody is linked to atherapeutic agent and is administered to a patient in need of suchtherapeutic agent. The therapeutic agent may be a toxin, if breasttissue needs to be selectively destroyed. This would be useful fortargeting and killing breast cancer cells. In another embodiment, thetherapeutic agent may be a growth or differentiation factor, which wouldbc useful for promoting breast cell function.

[0440] In another embodiment, an anti-BSP antibody may be linked to animaging agent that can be detected using, e.g., magnetic resonanceimaging, CT or PET. This would be useful for determining and monitoringbreast function, identifying breast cancer tumors, and identifyingnoncancerous breast diseases.

EXAMPLES Example 1

[0441] Gene Expression Analysis

[0442] BSGs were identified by a systematic analysis of gene expressiondata in the LIFESEQ® Gold database available from Incyte Genomics Inc(Palo Alto, Calif.) using the data mining software package CLASP™(Candidate Lead Automatic Search Program). CLASP™ is a set of algorithmsthat interrogate Incyte's database to identify genes that are bothspecific to particular tissue types as well as differentially expressedin tissues from patients with cancer. LifeSeq® Gold contains informationabout which genes are expressed in various tissues in the body and aboutthe dynamics of expression in both normal and diseased states. CLASP™first sorts the LifeSeq® Gold database into defined tissue types, suchas breast, ovary and prostate. CLASP™ categorizes each tissue sample bydisease state. Disease states include “healthy,” “cancer,” “associatedwith cancer,” “other disease” and “other.” Categorizing the diseasestates improves our ability to identify tissue and cancer-specificmolecular targets. CLASP™ then performs a simultaneous parallel searchfor genes that are expressed both (1) selectively in the defined tissuetype compared to other tissue types and (2) differentially in the“cancer” disease state compared to the other disease states affectingthe same, or different, tissues. This sorting is accomplished by usingmathematical and statistical filters that specify the minimum change inexpression levels and the minimum frequency that the differentialexpression pattern must be observed across the tissue samples for thegene to be considered statistically significant. The CLASPTM algorithmquantifies the relative abundance of a particular gene in each tissuetype and in each disease state.

[0443] To find the BSGs of this invention, the following specific CLASP™profiles were utilized: tissue-specific expression (CLASP 1), detectableexpression only in cancer tissue (CLASP 2), highest differentialexpression for a given cancer (CLASP 4); differential expression incancer tissue (CLASP 5), and. cDNA libraries were divided into 60 uniquetissue types (early versions of LifeSeq® had 48 tissue types). Genes orESTs were grouped into “gene bins,” where each bin is a cluster ofsequences grouped together where they share a common contig. Theexpression level for each gene bin was calculated for each tissue type.Differential expression significance was calculated with rigorousstatistical significant testing taking into account variations in samplesize and relative gene abundance in different libraries and within eachlibrary (for the equations used to determine statistically significantexpression see Audic and Claverie “The significance of digital geneexpression profiles,” Genome Res 7(10): 986-995 (1997), includingEquation 1 on page 987 and Equation 2 on page 988, the contents of whichare incorporated by reference). Differentially expressed tissue-specificgenes were selected based on the percentage abundance level in thetargeted tissue versus all the other tissues (tissue-specificity). Theexpression levels for each gene in libraries of normal tissues ornon-tumor tissues from cancer patients were compared with the expressionlevels in tissue libraries associated with tumor or disease(cancer-specificity). The results were analyzed for statisticalsignificance.

[0444] The selection of the target genes meeting the rigorous CLASP™profile criteria were as follows:

[0445] (a) CLASP 1: tissue-specific expression: To qualify as a CLASP 1candidate, a gene must exhibit statistically significant expression inthe tissue of interest compared to all other tissues. Only if the geneexhibits such differential expression with a 90% of confidence level isit selected as a CLASP 1 candidate.

[0446] (b) CLASP 2: detectable expression only in cancer tissue: Toqualify as a CLASP 2 candidate, a gene must exhibit detectableexpression in tumor tissues and undetectable expression in librariesfrom normal individuals and libraries from normal tissue obtained fromdiseased patients. In addition, such a gene must also exhibit furtherspecificity for the tumor tissues of interest.

[0447] (c) CLASP 5: differential expression in cancer tissue: To qualifyas a CLASP 5 candidate, a gene must be differentially expressed in tumorlibraries in the tissue of interest compared to normal libraries for alltissues. Only if the gene exhibits such differential expression with a90% of confidence level is it selected as a CLASP 5 candidate.

[0448] The CLASP™ scores for SEQ ID NO: 1-159 are listed below: SEQ IDNO: 1 DEX0249_1 CLASP2 SEQ ID NO: 2 DEX0249_2 CLASP2 SEQ ID NO: 3DEX0249_3 CLASP2 SEQ ID NO: 4 DEX0249_4 CLASP2 SEQ ID NO: 5 DEX0249_5CLASP2 SEQ ID NO: 6 DEX0249_6 CLASP2 SEQ ID NO: 7 DEX0249_7 CLASP2 SEQID NO: 8 DEX0249_8 CLASP2 SEQ ID NO: 9 DEX0249_9 CLASP2 SEQ ID NO: 11DEX0249_11 CLASP2 SEQ ID NO: 12 DEX0249_12 CLASP2 SEQ ID NO: 13DEX0249_13 CLASP2 SEQ ID NO: 14 DEX0249_14 CLASP2 SEQ ID NO: 15DEX0249_15 CLASP2 CLASP1 SEQ ID NO: 16 DEX0249_16 CLASP2 CLASP1 SEQ IDNO: 17 DEX0249_17 CLASP2 SEQ ID NO: 18 DEX0249_18 CLASP2 SEQ ID NO: 19DEX0249_19 CLASP2 SEQ ID NO: 20 DEX0249_20 CLASP2 SEQ ID NO: 21DEX0249_21 CLASP2 SEQ ID NO: 22 DEX0249_22 CLASP2 SEQ ID NO: 23DEX0249_23 CLASP2 SEQ ID NO: 24 DEX0249_24 CLASP2 SEQ ID NO: 25DEX0249_25 CLASP2 CLASP1 SEQ ID NO: 26 DEX0249_26 CLASP2 CLASP1 SEQ IDNO: 27 DEX0249_27 CLASP2 SEQ ID NO: 28 DEX0249_28 CLASP5 CLASP1 SEQ IDNO: 29 DEX0249_29 CLASP2 SEQ ID NO: 30 DEX0249_30 CLASP2 SEQ ID NO: 31DEX0249_31 CLASP2 SEQ ID NO: 32 DEX0249_32 CLASP2 SEQ ID NO: 33DEX0249_33 CLASP2 SEQ ID NO: 34 DEX0249_34 CLASP2 SEQ ID NO: 35DEX0249_35 CLASP2 SEQ ID NO: 36 DEX0249_36 CLASP2 SEQ ID NO: 37DEX0249_37 CLASP2 SEQ ID NO: 38 DEX0249_38 CLASP2 SEQ ID NO: 39DEX0249_39 CLASP2 SEQ ID NO: 40 DEX0249_40 CLASP2 SEQ ID NO: 41DEX0249_41 CLASP2 SEQ ID NO: 42 DEX0249_42 CLASP2 SEQ ID NO: 43DEX0249_43 CLASP2 SEQ ID NO: 44 DEX0249_44 CLASP2 SEQ ID NO: 45DEX0249_45 CLASP2 SEQ ID NO: 46 DEX0249_46 CLASP2 SEQ ID NO: 47DEX0249_47 CLASP2 SEQ ID NO: 48 DEX0249_48 CLASP2 SEQ ID NO: 49DEX0249_49 CLASP2 SEQ ID NO: 50 DEX0249_50 CLASP2 SEQ ID NO: 51DEX0249_51 CLASP2 SEQ ID NO: 52 DEX0249_52 CLASP2 SEQ ID NO: 53DEX0249_53 CLASP2 SEQ ID NO: 54 DEX0249_54 CLASP2 SEQ ID NO: 55DEX0249_55 CLASP2 SEQ ID NO: 56 DEX0249_56 CLASP2 SEQ ID NO: 57DEX0249_57 CLASP2 SEQ ID NO: 58 DEX0249_58 CLASP2 SEQ ID NO: 59DEX0249_59 CLASP2 SEQ ID NO: 60 DEX0249_60 CLASP2 SEQ ID NO: 61DEX0249_61 CLASP2 SEQ ID NO: 62 DEX0249_62 CLASP5 CLASP1 SEQ ID NO: 63DEX0249_63 CLASP5 CLASP1 SEQ ID NO: 64 DEX0249_64 CLASP5 CLASP1 SEQ IDNO: 65 DEX0249_65 CLASP5 CLASP1 SEQ ID NO: 66 DEX0249_66 CLASP2 SEQ IDNO: 67 DEX0249_67 CLASP2 SEQ ID NO: 68 DEX0249_68 CLASP5 CLASP1 SEQ IDNO: 69 DEX0249_69 CLASP2 SEQ ID NO: 70 DEX0249_70 CLASP2 SEQ ID NO: 71DEX0249_71 CLASP2 SEQ ID NO: 72 DEX0249_72 CLASP2 SEQ ID NO: 73DEX0249_73 CLASP2 SEQ ID NO: 74 DEX0249_74 CLASP2 SEQ ID NO: 75DEX0249_75 CLASP2 SEQ ID NO: 76 DEX0249_76 CLASP2 SEQ ID NO: 77DEX0249_77 CLASP2 SEQ ID NO: 78 DEX0249_78 CLASP5 CLASP1 SEQ ID NO: 79DEX0249_79 CLASP2 SEQ ID NO: 80 DEX0249_80 CLASP2 SEQ ID NO: 81DEX0249_81 CLASP2 CLASP1 SEQ ID NO: 82 DEX0249_82 CLASP2 SEQ ID NO: 83DEX0249_83 CLASP2 SEQ ID NO: 84 DEX0249_84 CLASP2 SEQ ID NO: 85DEX0249_85 CLASP2 SEQ ID NO: 86 DEX0249_86 CLASP2 SEQ ID NO: 87DEX0249_87 CLASP2 SEQ ID NO: 88 DEX0249_88 CLASP2 SEQ ID NO: 89DEX0249_89 CLASP2 SEQ ID NO: 90 DEX0249_90 CLASP2 CLASP1 SEQ ID NO: 91DEX0249_91 CLASP2 SEQ ID NO: 92 DEX0249_92 CLASP5 CLASP1 SEQ ID NO: 93DEX0249_93 CLASP2 SEQ ID NO: 94 DEX0249_94 CLASP2 SEQ ID NO: 95DEX0249_95 CLASP2 SEQ ID NO: 96 DEX0249_96 CLASP2 SEQ ID NO: 97DEX0249_97 CLASP2 SEQ ID NO: 98 DEX0249_98 CLASP2 SEQ ID NO: 99DEX0249_99 CLASP2 SEQ ID NO: 100 DEX0249_100 CLASP2 SEQ ID NO: 101DEX0249_101 CLASP2 SEQ ID NO: 102 DEX0249_102 CLASP2 SEQ ID NO: 103DEX0249_103 CLASP2 SEQ ID NO: 104 DEX0249_104 CLASP2 SEQ ID NO: 105DEX0249_105 CLASP5 CLASP1 SEQ ID NO: 106 DEX0249_106 CLASP5 CLASP1 SEQID NO: 107 DEX0249_107 CLASP5 CLASP1 SEQ ID NO: 108 DEX0249_108 CLASP5CLASP1 SEQ ID NO: 109 DEX0249_109 CLASP5 CLASP1 SEQ ID NO: 110DEX0249_110 CLASP2 SEQ ID NO: 111 DEX0249_111 CLASP2 SEQ ID NO: 112DEX0249_112 CLASP5 CLASP1 SEQ ID NO: 113 DEX0249_113 CLASP5 CLASP1 SEQID NO: 114 DEX0249_114 CLASP5 CLASP1 SEQ ID NO: 115 DEX0249_115 CLASP2SEQ ID NO: 116 DEX0249_116 CLASP2 SEQ ID NO: 117 DEX0249_117 CLASP2 SEQID NO: 118 DEX0249_118 CLASP2 SEQ ID NO: 119 DEX0249_119 CLASP2 SEQ IDNO: 120 DEX0249_120 CLASP2 SEQ ID NO: 121 DEX0249_121 CLASP2 SEQ ID NO:122 DEX0249_122 CLASP1 SEQ ID NO: 123 DEX0249_123 CLASP1 SEQ ID NO: 124DEX0249_124 CLASP5 CLASP1 SEQ ID NO: 125 DEX0249_125 CLASP5 CLASP1 SEQID NO: 126 DEX0249_126 CLASP2 SEQ ID NO: 127 DEX0249_127 CLASP2 SEQ IDNO: 128 DEX0249_128 CLASP2 SEQ ID NO: 129 DEX0249_129 CLASP5 CLASP1 SEQID NO: 130 DEX0249_130 CLASP5 CLASP1 SEQ ID NO: 131 DEX0249_131 CLASP5CLASP1 SEQ ID NO: 132 DEX0249_132 CLASP2 SEQ ID NO: 133 DEX0249_133CLASP2 SEQ ID NO: 134 DEX0249_134 CLASP2 SEQ ID NO: 135 DEX0249_135CLASP1 SEQ ID NO: 136 DEX0249_136 CLASP1 SEQ ID NO: 137 DEX0249_137CLASP1 SEQ ID NO: 138 DEX0249_138 CLASP2 CLASP1 SEQ ID NO: 139DEX0249_139 CLASP2 SEQ ID NO: 140 DEX0249_140 CLASP2 SEQ ID NO: 141DEX0249_141 CLASP2 SEQ ID NO: 142 DEX0249_142 CLASP2 SEQ ID NO: 143DEX0249_143 CLASP2 SEQ ID NO: 144 DEX0249_144 CLASP2 SEQ ID NO: 145DEX0249_145 CLASP2 SEQ ID NO: 146 DEX0249_146 CLASP2 SEQ ID NO: 147DEX0249_147 CLASP2 SEQ ID NO: 148 DEX0249_148 CLASP2 SEQ ID NO: 149DEX0249_149 CLASP2 SEQ ID NO: 150 DEX0249_150 CLASP2 SEQ ID NO: 151DEX0249_151 CLASP2 SEQ ID NO: 152 DEX0249_152 CLASP2 SEQ ID NO: 153DEX0249_153 CLASP2 SEQ ID NO: 154 DEX0249_154 CLASP2 SEQ ID NO: 155DEX0249_155 CLASP2 SEQ ID NO: 156 DEX0249_156 CLASP2 SEQ ID NO: 158DEX0249_158 CLASP2 SEQ ID NO: 159 DEX0249_159 CLASP2

Example 2

[0449] Relative Quantitation of Gene Expression

[0450] Real-Time quantitative PCR with fluorescent Taqman probes is aquantitation detection system utilizing the 5′-3′ nuclease activity ofTaq DNA polymerase. The method uses an internal fluorescentoligonucleotide probe (Taqman) labeled with a 5′ reporter dye and adownstream, 3′ quencher dye. During PCR, the 5′-3′ nuclease activity ofTaq DNA polymerase releases the reporter, whose fluorescence can then bedetected by the laser detector of the Model 7700 Sequence DetectionSystem (PE Applied Biosystems, Foster City, Calif., USA). Amplificationof an endogenous control is used to standardize the amount of sample RNAadded to the reaction and normalize for Reverse Transcriptase (RT)efficiency. Either cyclophilin, glyceraldehyde-3-phosphate dehydrogenase(GAPDH), ATPase, or 18S ribosomal RNA (rRNA) is used as this endogenouscontrol. To calculate relative quantitation between all the samplesstudied, the target RNA levels for one sample were used as the basis forcomparative results (calibrator). Quantitation relative to the“calibrator” can be obtained using the standard curve method or thecomparative method (User Bulletin #2: ABI PRISM 7700 Sequence DetectionSystem).

[0451] The tissue distribution and the level of the target gene areevaluated for every sample in normal and cancer tissues. Total RNA isextracted from normal tissues, cancer tissues, and from cancers and thecorresponding matched adjacent tissues. Subsequently, first strand cDNAis prepared with reverse transcriptase and the polymerase chain reactionis done using primers and Taqman probes specific to each target gene.The results are analyzed using the ABI PRISM 7700 Sequence Detector. Theabsolute numbers are relative levels of expression of the target gene ina particular tissue compared to the calibrator tissue.

[0452] One of ordinary skill can design appropriate primers. Therelative levels of expression of the BSNA versus normal tissues andother cancer tissues can then be determined. All the values are comparedto normal thymus (calibrator). These RNA samples are commerciallyavailable pools, originated by pooling samples of a particular tissuefrom different individuals.

[0453] The relative levels of expression of the BSNA in pairs ofmatching samples and 1 cancer and 1 normal/normal adjacent of tissue mayalso be determined. All the values are compared to normal thymus(calibrator). A matching pair is fonned by mRNA from the cancer samplefor a particular tissue and mRNA from the normal adjacent sample forthat same tissue from the same individual.

[0454] In the analysis of matching samples, the BSNAs that show a highdegree of tissue specificity for the tissue of interest. These resultsconfirm the tissue specificity results obtained with normal pooledsamples.

[0455] Further, the level of mRNA expression in cancer samples and theisogenic normal adjacent tissue from the same individual are compared.This comparison provides an indication of specificity for the cancerstage (e.g. higher levels of mRNA expression in the cancer samplecompared to the normal adjacent).

[0456] Altogether, the high level of tissue specificity, plus the mRNAoverexpression in matching samples tested are indicative of SEQ ID NO: 1through 159 being a diagnostic marker for cancer.

[0457] DEX0093_(—)11 (mam029-sqmam036) Sequence Sequence ID NoDex0093_11 DEX0249_15 (SEQ ID NO:15) (mam029-sqmam036) DEX0249_16 (SEQID NO:16)

[0458] Semi-quantitative PCR was done using the following primers:Primer DexSeqID From To Primer Length sqmam036F DEX0249_16 395 417 23sqmam036F DEX0249_15  50  72 23 sqmam036R DEX0249_16 684 661 24sqmam036R DEX0249_15 339 316 24

[0459] Data from the semiQ-PCR experiment showed that sqmam036 wasexpressed in normal breast in higher levels compared with other 11normal tissue samples tested, and expressed in breast carcinoma whileabsent in other 11 cancer tissue types (bladder, colon, kidney, lung,ovary, liver, pancreas, prostate, stomach, testis and uterus). Sqmam123was advanced to quantitative PCR and named mam029.

[0460] Quantitative PCR was done using the following primers: PrimerDexSeqID From To Primer Length mam029F DEX0249_16 859 876 18 mam029FDEX0249_15 514 531 18 mam029R DEX0249_16 992 970 23 mam029R DEX0249_15647 625 23 mam029 probe DEX0249_16 961 934 28 mam029 probe DEX0249_15616 589 28

[0461] The relative levels of expression of mam029 in 36 normal samplesfrom 25 different tissues were measured. All the values are compared tonormal testis (calibrator). These RNA samples are commercially availablepools, originated by pooling samples of a particular tissue fromdifferent individuals; except for the blood samples that they are normalsamples from a single individual. Tissue Normal Adrenal Gland 0.0018Bladder 0.0000 Brain 0.0013 Cervix 0.0154 Colon 0.0000 Endometrium0.0000 Esophagus 0.0016 Heart 0.0000 Kidney 0.0000 Liver 0.0000 Lung0.0004 Mammary gland 3.1167 Muscle 0.0041 Ovary 0.00 Pancreas 0.00Prostate 0.00 Rectum 0.00 Small 0.00 Intestine Spleen 0.00 Stomach 0.00Testis 1.00 Thymus 0.00 Trachea 0.00 Uterus 0.00 Blood B1 0.49 Blood B100.00 Blood B11 0.00 Blood B12 0.00 Blood B13 0.00 Blood B14 0.00 BloodB15 0.00 Blood B4 0.00 Blood B5 0.00 Blood B6 0.00 Blood B7 0.00 BloodB8 0.00

[0462] The relative levels of expression in Table 1 show that mam029mRNA is highly expressed in the pool of normal mammary gland comparedwith the other normal tissue analyzed.

[0463] The absolute numbers in Table 1 were obtained analyzing pools ofsamples of a particular tissue from different individuals. They cannotbe compared to the absolute numbers originated from RNA obtained fromtissue samples of a single individual in Table 2.

[0464] The relative levels of expression of mam029 in 48 pairs ofmatching samples were measured. All the values are compared to normaltestis (calibrator). A matching pair is formed by mRNA from the cancersample for a particular tissue and mRNA from the normal adjacent samplefor that same tissue from the same individual. In addition, 1 unmatchedcancer sample (from ovary) and 1 unmatched normal sample (from ovary)were also tested. Normal Sample Adjacent ID Tissue Cancer Tissue NormalMam497M mammary gland 1 34.66 7.36 Mam173M mammary gland 2 0.26 25.28Mam726M mammary gland 3 26.45 7.44 MamS516 mammary gland 4 0.48 0.22MamS621 mammary gland 5 7.70 0.47 MamS079 mammary gland 6 0.92 1.32Mam517 mammary gland 7 0.40 0.83 Mam19DN mammary gland 8 27.76 16.97Mam522 mammary gland 9 13.09 1.39 MamB011X mammary gland 10 0.02 4.77MamS127 mammary gland 11 0.00 0.63 Mam51DN mammary gland 12 1.79 4.69Mam220 mammary gland 13 5.37 18.25 Mam245M mammary gland 14 0.02 1.72Mam162X mammary gland 15 5.31 0.15 MamS123 mammary gland 16 0.06 2.35MamS997 mammary gland 17 0.52 1.04 Mam543M mammary gland 18 0.00 0.00Mam976M mammary gland 19 14.47 1.64 Mam76DN mammary gland 20 28.84 44.63Mam699F mammary gland 21 0.03 0.56 Mam42DN mammary gland 22 0.17 2.46MamS570 mammary gland 23 0.04 19.23 MamS918 mammary gland 24 24.50 12.34MamS854 mammary gland 25 3.19 0.76 Mam986 mammary gland 26 9.71 5.78MamS967 mammary gland 27 45.10 2.66 Mam355 mammary gland 28 56.69 0.10MamS699 mammary gland 29 0.30 1.89 MamA06X mammary gland 30 121.1 1.25Bld32XK bladder 1 0.01 0.00 Bld66X bladder 2 0.00 0.00 BldTR17 bladder 30.00 0.00 Bld46XK bladder 4 0.00 0.00 BldTR14 bladder 5 0.00 0.00 ClnB56colon 1 0.00 0.00 ClnDC63 colon 2 0.007366 0.00 CvxKS52 cervix 1 0.000.00 CvxNK24 cervix 2 0.00 0.00 CvxKS83 cervix 3 0.00 0.00 CvxNK23cervix 4 0.00 0.00 Endo10479 endometrium 1 0.00 0.07 Endo12XAendometrium 2 0.00 0.00 Endo5XA endometrium 3 0.00 0.00 Endo65RAendometrium 4 0.00 0.00 Endo28XA endometrium 5 0.00 0.00 Endo3AXendometrium 6 0.52 0.00 Kid98XD kidney 1 0.00 0.00 Kid6XD kidney 2 0.000.00 Kid710K kidney 3 0.00 0.00 Liv175L liver 1 0.00 0.00 Liv187L liver2 0.00 0.00 Liv15XA liver 3 0.07 0.00 Lng47XQ lung 1 0.08 0.00 LngAC88lung 2 0.00 0.00 LngAC90 lung 3 0.00 0.00 LngSQ80 lung 4 0.00 0.00Ovr1118 ovary 1 0.00 Ovr32RA ovary 2 0.00 Pan77X pancreas 1 0.00 0.00Pan82XP pancreas 2 0.01 0.00 Pro109XB prostate 0.00 0.08 Skn248S skin 10.00 0.04 Skn287S skin 2 0.00 0.00 SmIntH89 small intestine 1 0.00 0.00SmInt21XA small intestine 2 0.01 0.00 Sto115S stomach 1 0.00 0.00 Sto15Sstomach 2 0.00 0.00 StoMT54 stomach 3 0.00 0.00 Thr590D thymus 1 0.000.00 Tst647T testis 1 0.00 0.09 Utr141XO uterus 1 0.01 0.00 Utr23XUuterus 2 0.00 0.00 Utr85XU uterus 3 0.05 0.00 Utr135XO uterus 4 0.000.00

[0465] The table above represents 148 samples in 17 different tissues.Table 1 and Table 2 represent a combined total of 184 samples in 27human tissue types. Comparisons of the level of mRNA expression inbreast cancer samples and the normal adjacent tissue from the sameindividuals are shown in Table 2. Mam029 is expressed at higher levelsin 14 of 30 (47%) cancer samples (mammary gland 1, 3-5, 8, 9, 15, 19,24-28, 30) compared to normal adjacent tissue.

[0466] Mam029 is highly specific for breast, showing no expression inmost of the samples analyzed for other cancers (specificity: 98%;specificity was calculated as the percentage of samples other thanbreast with level of expression below of 1/10 of the median for breastcancer samples).

[0467] Northern blot analysis for mam029 revealed a 1.2 kb transcript.

[0468] DEX0093_(—)35 Sqmam046 Sequence Sequence ID # Dex0093_35(sqmam046) DEX0249_50 (SEQ ID NO:50)

[0469] Semi-quantitative PCR was done using the following primers:Primer DexSeqID From To Primer Length sqmam046F DEX0249_50 3 24 22sqmam046R DEX0249_50 380 361 20

[0470] The relative levels of expression of sqmam046 in 12 normalsamples from 12 different tissues are shown below. These RNA samples arefrom single individual or are commercially available pools, originatedby pooling samples of a particular tissue from different individuals.Using Polymerase Chain Reaction (PCR) technology expression levels wereanalyzed from four 10×serial cDNA dilutions in duplicate. Relativeexpression levels of 0, 1, 10, 100 and 1000 are used to evaluate geneexpression. A positive reaction in the most dilute sample indicates thehighest relative expression value. TISSUE NORMAL Breast 1 Colon 10Endometrium 10 Kidney 0 Liver 0 Lung 1 Ovary 1 Prostate 100 SmallIntestine 1 Stomach 100 Testis 1 Uterus 0

[0471] Relative levels of expression in the table above show that normalprostate and stomach exhibit the highest expression of sqmam046,followed by colon and endometrium.

[0472] The relative levels of expression of sqmam046 in 12 cancersamples from 12 different tissues are shown below. Using PolymeraseChain Reaction (PCR) technology expression levels were analyzed fromfour 10×serial cDNA dilutions in duplicate. Relative expression levelsof 0, 1, 10, 100 and 1000 are used to evaluate gene expression. Apositive reaction in the most dilute sample indicates the highestrelative expression value. TISSUE CANCER bladder 1 breast 100 colon 1kidney 0 liver 0 lung 0 ovary 0 pancreas 0 prostate 10 stomach 0 testes0 Uterus 1

[0473] Relative levels of expression in the table above shows thatsqmam046 is highly expressed in breast carcinoma when compared with theother cancer samples tested.

[0474] The relative levels of expression of sqmam046 in 6 mammary glandcancer matching samples are shown below. A matching pair is formed bymRNA from the cancer sample for a particular tissue and mRNA from thenormal adjacent sample for that same tissue from the same individual.

[0475] Using Polymerase Chain Reaction (PCR) technology expressionlevels were analyzed from four 10×serial cDNA dilutions in duplicate.Relative expression levels of 0, 1, 10, 100 and 1000 are used toevaluate gene expression. A positive reaction in the most dilute sampleindicates the highest relative expression value. NORMAL ADJACENT SAMPLEID TISSUE CANCER TISSUE S99522A/B mammary gland 1 10 0 4005724A2/B3mammary gland 2 1 1 4005599A4/B2 mammary gland 3 1 0 4005629A2/B2mammary gland 4 10 1 S9822245A/B mammary gland 5 100 10 S9819997A/Bmammary gland 6 1 1

[0476] Relative levels of expression in the Table above shows thatsqmam046 is upregulated in 4 out of 6 (67%) of the matching samplesanalyzed. Experiments are underway to design and test primers and probefor quantitative PCR.

[0477] DEX0093_(—)46

[0478] Sqmam050 Sequence Sequence ID # Dex0093_46 (sqmam050) DEX0249_69(SEQ ID NO:69) DEX0249_70 (SEQ ID NO:70)

[0479] Semi-quantitative PCR was done using the following primers:Primer DexSeqID From To Primer Length sqmam050F DEX0249_70 68 92 25sqmam050F DEX0249_69 68 92 25 sqmam050R DEX0249_70 523 502 22 sqmam050RDEX0249_69 523 502 22

[0480] The relative levels of expression of sqmam050 in 12 normalsamples from 12 different tissues are shown below. These RNA samples arefrom single individual or are commercially available pools, originatedby pooling samples of a particular tissue from different individuals.Using Polymerase Chain Reaction (PCR) technology expression levels wereanalyzed from four 10×serial cDNA dilutions in duplicate. Relativeexpression levels of 0, 1, 10, 100 and 1000 are used to evaluate geneexpression. A positive reaction in the most dilute sample indicates thehighest relative expression value. TISSUE NORMAL Breast 0 Colon 0Endometrium 0 Kidney 0 Liver 0 Lung 0 Ovary 0 Prostate 0 Small Intestine0 Stomach 0 Testis 0 Uterus 0

[0481] Relative levels of expression in Table 1 show that none of thenormal tissues tested show expression of sqmam050.

[0482] The relative levels of expression of sqmam050 in 12 cancersamples from 12 different tissues are shown below. Using PolymeraseChain Reaction (PCR) technology expression levels were analyzed fromfour 10×serial cDNA dilutions in duplicate. Relative expression levelsof 0, 1, 10, 100 and 1000 are used to evaluate gene expression. Apositive reaction in the most dilute sample indicates the highestrelative expression value. TISSUE CANCER bladder 1 breast 0 colon 1kidney 1 liver 0 lung 1 ovary 0 pancreas 1 prostate 10 stomach 0 testes1 uterus 10

[0483] Relative levels of expression in Table 2 show that sqmam050 isexpressed in the cancer samples tested, with the highest expression inprostate and uterus.

[0484] The relative levels of expression of sqmam050 in 6 mammary glandcancer matching samples are show below. A matching pair is formed bymRNA from the cancer sample for a particular tissue and mRNA from thenormal adjacent sample for that same tissue from the same individual.

[0485] Using Polymerase Chain Reaction (PCR) technology expressionlevels were analyzed from four 10×serial cDNA dilutions in duplicate.Relative expression levels of 0, 1, 10, 100 and 1000 are used toevaluate gene expression. A positive reaction in the most dilute sampleindicates the highest relative expression value. NORMAL SAMPLE ADJACENTID TISSUE CANCER TISSUE S99522A/B mammary gland 1 10 1 4005724A2/B3mammary gland 2 1 0 4005599A4/B2 mammary gland 3 1 0 4005629A2/B2mammary gland 4 1 1 S9822245A/B mammary gland 5 0 0 S9819997A/B mammarygland 6 10 1

[0486] Relative levels of expression in Table 2 shows that sqmam050 isupregulated in 4 out of 6 (67%) of the matching samples analyzed.

[0487] Experiments are underway to design and test primers and probe forquantitative PCR.

[0488] DEX0093_(—)65

[0489] Sqmam054 Sequence Sequence ID # Dex0093_65 (sqmam054) DEX0249_93(SEQ ID NO:93) DEX0249_94 (SEQ ID NO:94)

[0490] Semi-quantitative PCR was done using the following primers:Primer DexSeqID From To Primer Length sqmam054F DEX0249_94 115 132 18sqmam054F DEX0249_93 117 134 18 sqmam054R DEX0249_94 251 234 18sqmam054R DEX0249_93 253 236 18

[0491] Table 1. The absolute numbers are relative levels of expressionof sqmam054 in 12 normal samples from 12 different tissues. These RNAsamples are from single individual or are commercially available pools,originated by pooling samples of a particular tissue from differentindividuals.. Using Polymerase Chain Reaction (PCR) technologyexpression levels were analyzed from four 10×serial cDNA dilutions induplicate. Relative expression levels of 0, 1, 10, 100 and 1000 are usedto evaluate gene expression. A positive reaction in the most dilutesample indicates the highest relative expression value. TISSUE NORMALBreast 0 Colon 0 Endometrium 0 Kidney 0 Liver 0 Lung 0 Ovary 0 Prostate0 Small Intestine 0 Stomach 1 Testis 0 Uterus 1

[0492] Relative levels of expression in Table 1 show expression ofsqmam054 in stomach and uterus normal tissues.

[0493] Table 2. The relative levels of expression of sqmam054 in 12cancer samples from 12 different tissues are shown below. UsingPolymerase Chain Reaction (PCR) technology expression levels wereanalyzed from four 10×serial cDNA dilutions in duplicate. Relativeexpression levels of 0, 1, 10, 100 and 1000 are used to evaluate geneexpression. A positive reaction in the most dilute sample indicates thehighest relative expression value. TISSUE CANCER bladder 1 breast 1colon 0 kidney 1 liver 0 lung 1 ovary 0 pancreas 1 prostate 1 stomach 10Testes 1 Uterus 1

[0494] Relative levels of expression in Table 2 show that sqmam054 isexpressed in some of the cancer samples tested. The highest level ofexpression in stomach cancer tissue.

[0495] Table 3. The absolute numbers are relative levels of expressionof sqmam054 in 6 mammary gland cancer matching samples. A matching pairis formed by mRNA from the cancer sample for a particular tissue andmRNA from the normal adjacent sample for that same tissue from the sameindividual.

[0496] Using Polymerase Chain Reaction (PCR) technology expressionlevels were analyzed from four 10×serial cDNA dilutions in duplicate.Relative expression levels of 0, 1, 10, 100 and 1000 are used toevaluate gene expression. A positive reaction in the most dilute sampleindicates the highest relative expression value. NORMAL ADJACENT SAMPLEID TISSUE CANCER TISSUE S99522A/B mammary gland 1 1 0 4005724A2/B3mammary gland 2 0 0 4005599A4/B2 mammary gland 3 1 0 4005629A2/B2mammary gland 4 1 0 S9822245A/B mammary gland 5 0 10 S9819997A/B mammarygland 6 0 0

[0497] Relative levels of expression in Table 2 shows that sqmam054 isupregulated in 3 out of 6 (50%) of the matching samples analyzed, andtwo matching samples do not express sqmam054 in any of both samples:cancer and NAT.

[0498] Experiments are underway to design and test primers and probe forquantitative PCR.

Example 2B

[0499] Custom Microarray Experiment

[0500] Custom oligonucleotide microarrays were provided by AgilentTechnologies, Inc. (Palo Alto, Calif.). The microarrays were fabricatedby Agilent using their technology for the in-situ synthesis of 60meroligonucleotides (Hughes, et al. 2001, Nature Biotechnology 19:342-347).The 60mer microarray probes were designed by Agilent, from genesequences provided by diaDexus, using Agilent proprietary algorithms.Whenever possible two different 60mers were designed for each gene ofinterest.

[0501] All microarray experiments were two-color experiments and werepreformed using Agilent-recommended protocols and reagents. Briefly,each microarray was hybridized with cRNAs synthesized from polyA+ RNA,isolated from cancer and normal tissues, labeled with fluorescent dyesCyanine3 and Cyanine5 (NEN Life Science Products, Inc., Boston, Mass.)using a linear amplification method (Agilent). In each experiment theexperimental sample was polyA+RNA isolated from cancer tissue from asingle individual and the reference sample was a pool of polyA+ RNAisolated from normal tissues of the same organ as the cancerous tissue(i.e. normal breast tissue in experiments with breast cancer samples).Hybridizations were carried out at 60° C., overnight using Agilentin-situ hybridization buffer. Following washing, arrays were scannedwith a GenePix 4000B Microarray Scanner (Axon Instruments, Inc., UnionCity, Calif.). The resulting images were analyzed with GenePix Pro 3.0Microarray Acquisition and Analysis Software (Axon). A total of 20experiments comparing the expression patterns of breast cancer derivedpolyA+ RNA (6 stage 1 cancers, 12 stage 12 cancers, 2 stage 3 cancers)to polyA+ RNA isolated from a pool of 10 normal breast tissues wereanalyzed.

[0502] Data normalization and expression profiling were done withExpressionist software from GeneData Inc. (Daly City, Calif./Basel,Switzerland). Gene expression analysis was performed using onlyexperiments that meet certain quality criteria. The quality criteriathat experiments must meet are a combination of evaluations performed bythe Expressionist software and evaluations performed manually using rawand normalized data. To evaluate raw data quality, detection limits (themean signal for a replicated negative control+2 Standard Deviations(SD)) for each channel were calculated. The detection limit is a measureof non-specific hybridization. Arrays with poor detection limits werenot analyzed and the experiments were repeated. To evaluate normalizeddata quality, positive control elements included in the array wereutilized. These array features should have a mean ratio of 1 (nodifferential expression). If these features have a mean ratio of greaterthan 1.5-fold up or down, the experiments were not analyzed further andwere repeated. In addition to traditional scatter plots demonstratingthe distribution of signal in each experiment, the Expressionistsoftware also has minimum thresholding criteria that employ user definedparameters to identify quality data. Only those features that meet thethreshold criteria were included in the filtering and analyses carriedout by Expressionist. The thresholding settings employed require aminimum area percentage of 60% [(% pixels>background+2SD)−(% pixelssaturated)], and a minimum signal to noise ratio of 2.0 in bothchannels. By these criteria, very low expressors and saturated featureswere not included in analysis.

[0503] Relative expression data was collected from Expressionist basedon filtering and clustering analyses. Up- and down-regulated genes wereidentified using criteria for percentage of valid values obtained, andthe percentage of experiments in which the gene is up- ordown-regulated. These criteria were set independently for each data set,depending on the size and the nature of the data set. The results forthe statistically significant upregulated and downregulated genes areshown in Table 1. The first three columns of the table containinformation about the sequence itself (Oligo ID, Parent ID, andPatent#), the next 3 columns show the results obtained. ‘% valid’indicates the percentage of 20 unique experiments total in which a validexpression value was obtained, ‘% up’ indicates the percentage of 20experiments in which up-regulation of at least 2.5-fold was observed,and ‘% down’ indicates the percentage of the 20 experiments in whichdown-regulation of at least 2.5-fold was observed. The last column inTable 1 describes the location of the microarray probe (oligo) relativeto the parent sequence. Additional sequences were examined but the datawere inconclusive. Sensitivity data for DEX0093 series microarrayfeatures. Sensitivity of up Oligo Parent and down regulation Seq OligoIDID Patent # %valid %up %down location 20173 4795 DEX0093_86 65% 40% 5%653-712 DEX0249_124

Example 3

[0504] Protein Expression

[0505] The BSNA is amplified by polymerase chain reaction (PCR) and theamplified DNA fragment encoding the BSNA is subcloned in pET-21d forexpression in E. coli. In addition to the BSNA coding sequence, codonsfor two amino acids, Met-Ala, flanking the NH₂-terminus of the codingsequence of BSNA, and six histidines, flanking the COOH-terminus of thecoding sequence of BSNA, are incorporated to serve as initiatingMet/restriction site and purification tag, respectively.

[0506] An over-expressed protein band of the appropriate molecularweight may be observed on a Coomassie blue stained polyacrylamide gel.This protein band is confirmed by Western blot analysis using monoclonalantibody against 6×Histidine tag.

[0507] Large-scale purification of BSP was achieved using cell pastegenerated from 6-liter bacterial cultures, and purified usingimmobilized metal affinity chromatography (IMAC). Soluble fractions thathad been separated from total cell lysate were incubated with a nicklechelating resin. The column was packed and washed with five columnvolumes of wash buffer. BSP was eluted stepwise with variousconcentration imidazole buffers.

Example 4

[0508] Protein Fusions

[0509] Briefly, the human Fc portion of the IgG molecule can be PCRamplified, using primers that span the 5′ and 3′ ends of the sequencedescribed below. These primers also should have convenient restrictionenzyme sites that will facilitate cloning into an expression vector,preferably a mammalian expression vector. For example, if pC4 (AccessionNo. 209646) is used, the human Fc portion can be ligated into the BamHIcloning site. Note that the 3′ BamHI site should be destroyed. Next, thevector containing the human Fc portion is re-restricted with BamHI,linearizing the vector, and a polynucleotide of the present invention,isolated by the PCR protocol described in Example 2, is ligated intothis BamHI site. Note that the polynucleotide is cloned without a stopcodon, otherwise a fusion protein will not be produced. If the naturallyoccurring signal sequence is used to produce the secreted protein, pC4does not need a second signal peptide. Alternatively, if the naturallyoccurring signal sequence is not used, the vector can be modified toinclude a heterologous signal sequence. See, e.g., WO 96/34891.

Example 5

[0510] Production of an Antibody from a Polypeptide

[0511] In general, such procedures involve immunizing an animal(preferably a mouse) with polypeptide or, more preferably, with asecreted polypeptide-expressing cell. Such cells may be cultured in anysuitable tissue culture medium; however, it is preferable to culturecells in Earle's modified Eagle's medium supplemented with 10% fetalbovine serum (inactivated at about 56° C.), and supplemented with about10 g/l of nonessential amino acids, about 1,000 U/ml of penicillin, andabout 100, μg/ml of streptomycin. The splenocytes of such mice areextracted and fused with a suitable myeloma cell line. Any suitablemyeloma cell line may be employed in accordance with the presentinvention; however, it is preferable to employ the parent myeloma cellline (SP20), available from the ATCC. After fusion, the resultinghybridoma cells are selectively maintained in HAT medium, and thencloned by limiting dilution as described by Wands et al.,Gastroenterology 80: 225-232 (1981).

[0512] The hybridoma cells obtained through such a selection are thenassayed to identify clones which secrete antibodies capable of bindingthe polypeptide. Alternatively, additional antibodies capable of bindingto the polypeptide can be produced in a two-step procedure usinganti-idiotypic antibodies. Such a method makes use of the fact thatantibodies are themselves antigens, and therefore, it is possible toobtain an antibody which binds to a second antibody. In accordance withthis method, protein specific antibodies are used to immunize an animal,preferably a mouse. The splenocytes of such an animal are then used toproduce hybridoma cells, and the hybridoma cells are screcned toidentify clones which produce an antibody whose ability to bind to theprotein-specific antibody can be blocked by the polypeptide. Suchantibodies comprise anti-idiotypic antibodies to the protein specificantibody and can be used to immunize an animal to induce formation offurther protein-specific antibodies. Using the Jameson-Wolf methods thefollowing epitopes were predicted. (Jameson and Wolf, CABIOS, 4(1),181-186, 1988, the contents of which are incorporated by reference).

[0513] Examples of post-translational modifications (PTMs) of the BSPsof this invention are listed below. In addition, antibodies thatspecifically bind such post-translational modifications may be useful asa diagnostic or as therapeutic. Using the ProSite database (Bairoch etal., Nucleic Acids Res. 25(1):217-221 (1997), the contents of which areincorporated by reference), the following PTMs were predicted for theBSPs of the invention(http://npsa-pbil.ibcp.fr/cgi-bin/npsa_automat.pl?page=npsa_prosite.htmlmost recently accessed Oct. 23, 2001). For full definitions of the PTMssee http://www.expasy.org/cgi-bin/prosite-list.pl most recently accessedOct. 23, 2001. DEX0249_160 Asn_Glycosylation 2-5; Ck2_Phospho_Site 4-7;DEX0249_161 Pkc_Phospho_Site 11-13; DEX0249_162 Myristyl 2-7;DEX0249_163 Ck2_Phospho_Site 2-5;42-45; DEX0249_164 Myristyl 16-21;Pkc_Phospho_Site 20-22; DEX0249_165 Ck2_Phospho_Site 26-29; DEX0249_166Asn_Glycosylation 4-7; DEX0249_167 Asn_Glycosylation 9-12; DEX0249_168Ck2_Phospho_Site 48-51; Pkc_Phospho_Site 59-61; DEX0249_169Ck2_Phospho_Site 81-84; Myristyl 17-22;26-31;42-47; Pkc_Phospho_Site10-12;83-85; DEX0249_170 Ck2_Phospho_Site 3-6; Pkc_Phospho_Site 5-7;DEX0249_171 Camp_Phospho_Site 69-72; Ck2_Phospho_Site 20-23; Myristyl13-18; Pkc_Phospho_Site 20-22;54-56;63-65;68-70; DEX0249_172Pkc_Phospho_Site 25-27; DEX0249_173 Glycosaminoglycan 25-28;Pkc_Phospho_Site 25-27;42-44; DEX0249_174 Myristyl 20-25;Pkc_Phospho_Site 6-8; DEX0249_175 Asn_Glycosylation 2-5; Myristyl 16-21;DEX0249_176 Pkc_Phospho_Site 8-10; DEX0249_177 Ck2_Phospho_Site 4-7;DEX0249_178 Pkc_Phospho_Site 8-10; DEX0249_179 Myristyl 34-39;Pkc_Phospho_Site 51-53; DEX0249_180 Ck2_Phospho_Site 19-22;21-24;Myristyl 13-18;38-43; DEX0249_182 Ck2_Phospho_Site 3-6;39-42;Pkc_Phospho_Site 3-5;39-41; DEX0249_183 Ck2_Phospho_Site 39-42; Myristyl17-22; Pkc_Phospho_Site 53- 55; DEX0249_184 Myristyl 18-23; DEX0249_187Myristyl 39-44; DEX0249_188 Myristyl 13-18;52-57;68-73; Pkc_Phospho_Site32-34;56-58;64- 66;65-67; DEX0249_189 Myristyl 17-22; Pkc_Phospho_Site22-24; DEX0249_190 Amidation 6-9; Ck2_Phospho_Site 27-30;Tyr_Phospho_Site 8- 16;9-16; DEX0249_192 Ck2_Phospho_Site 23-26;Pkc_Phospho_Site 23-25; DEX0249_193 Amidation 217-220; Camp_Phospho_Site84-87; Ck2_Phospho_Site 20-23;22-25;31-34; Myristyl 18-23;42-47;57-62;58-63;73-78;135-140;175-180;201-206;204-209;285-290; Pkc_Phospho_Site139-141;286-288;289-291; DEX0249_194 Pkc_Phospho_Site 26-28; DEX0249_195Myristyl 16-21; Pkc_Phospho_Site 36-38;70-72; DEX0249_196Camp_Phospho_Site 74-77; Ck2_Phospho_Site 79-82; Myristyl 12-17;Pkc_Phospho_Site 16-18;30-32;73-75;79-81; DEX0249_197 Asn_Glycosylation203-206;243-246;485-488;548-551;594-597; Ck2_Phospho_Site176-179;337-340;345-348;389-392;398-401;407-410;431-434;452-455;465-468;476-479;503-506;550- 553;563-566;Myristyl 520-525;538-543;558-563; Pkc_Phospho_Site45-47;176-178;195-197;205-207;217-219;221-223;232-234;312-314;315-317;320-322;362-364;365-367;413-415;431-433;530-532;614-616;633-635; Prokar_Lipoprotein 133- 143;DEX0249_198 Amidation 7-10; Pkc_Phospho_Site 5-7; DEX0249_199Pkc_Phospho_Site 3-5; DEX0249_200 Myristyl 32-37; DEX0249_202Ck2_Phospho_Site 49-52;53-56; DEX0249_204 Asn_Glycosylation 146-149;Ck2_Phospho_Site 19-22;43-46;73- 76;120-123; Pkc_Phospho_Site50-52;127-129;131-133; DEX0249_205 Ck2_Phospho_Site 3-6;85-88; Myristyl108-113; Pkc_Phospho_Site 44-46;93-95; DEX0249_206 Pkc_Phospho_Site12-14;20-22;23-25; DEX0249_207 Ck2_Phospho_Site 103-106; Myristyl46-51;71-76; Pkc_Phospho_Site 39-41;121-123; DEX0249_208Asn_Glycosylation 37-40; DEX0249_209 Ck2_Phospho_Site 7-10;23-26;28-31;Pkc_Phospho_Site 22-24; DEX0249_210 Myristyl 55-60; Pkc_Phospho_Site21-23; Tyr_Phospho_Site 50- 57; DEX0249_212 Pkc_Phospho_Site 23-25;DEX0249_213 Pkc_Phospho_Site 22-24; DEX0249_215 Asn_Glycosylation80-83;127-130; Ck2_Phospho_Site 18-21; Myristyl2-7;56-61;71-76;86-91;116-121;120-125; Pkc_Phospho_Site 75-77;Prokar_Lipoprotein 9-19; DEX0249_216 Ck2_Phospho_Site 41-44; DEX0249_217Pkc_Phospho_Site 57-59; DEX0249_218 Leucine_Zipper 22-43;Pkc_Phospho_Site 42-44;59-61; Prokar_Lipoprotein 50-60; Tyr_Phospho_Site61-68; DEX0249_219 Pkc_Phospho_Site 8-10; DEX0249_220 Asn_Glycosylation18-21; Pkc_Phospho_Site 20-22; DEX0249_222 Ck2_Phospho_Site 9-12;22-25;Pkc_Phospho_Site 19-21; Tyr_Phospho_Site 21-28; DEX0249_223Ck2_Phospho_Site 4-7; Myristyl 12-17;21-26; Pkc_Phospho_Site 4-6;DEX0249_224 Myristyl 31-36; Pkc_Phospho_Site 42-44; DEX0249_225Pkc_Phospho_Site 19-21; DEX0249_227 Ck2_Phospho_Site 29-32; Myristyl40-45; DEX0249_228 Ck2_Phospho_Site 8-11;14-17; Pkc_Phospho_Site22-24;76-78; DEX0249_229 Ck2_Phospho_Site 68-71; Myristyl 6-11;50-55;Pkc_Phospho_Site 62-64; DEX0249_230 Myristyl 5-10; Pkc_Phospho_Site21-23; DEX0249_231 Asn_Glycosylation 39-42; Pkc_Phospho_Site 43-45;DEX0249_233 Myristyl 25-30; DEX0249_234 Amidation 145-148;Ck2_Phospho_Site 29-32;30-33;56-59;96- 99;98-101; Myristyl85-90;92-97;94-99; Pkc_Phospho_Site 56- 58;63-65;103-105; DEX0249_235Asn_Glycosylation 56-59;171-174;399-402; Camp_Phospho_Site51-54;873-876; Ck2_Phospho_Site 13-16;123-126;182-185;222-225;266-269;320-323;649-652;682-685;795-798;832-835;845- 848;892-895;Myristyl 108-113;315-320;468-473;499-504;581- 586; Pkc_Phospho_Site60-62;101-103;146-148;163-165;166-168;222-224;279-281;289-291;336-338;337-339;391-393;574-576;607-609;655-657;673-675;784-786;809-811;832-834;856-858;920-922;932-934; Tyr_Phospho_Site 431-438; DEX0249_236Pkc_Phospho_Site 47-49; DEX0249_237 Ck2_Phospho_Site 23-26;31-34;DEX0249_238 Myristyl 20-25; DEX0249_239 Camp_Phospho_Site 11-14;Pkc_Phospho_Site 9-11; DEX0249_240 Camp_Phospho_Site 34-37;Helix_Loop_Helix 13-28; DEX0249_241 Ck2_Phospho_Site 20-23;34-37;Myristyl 3-8; DEX0249_243 Asn_Glycosylation 26-29;33-36;Ck2_Phospho_Site 35-38; Pkc_Phospho_Site 16-18; DEX0249_244 Myristyl13-18; Pkc_Phospho_Site 52-54; DEX0249_245 Ck2_Phospho_Site 17-20;Pkc_Phospho_Site 11-13;17-19; DEX0249_246 Amidation 10-13;Asn_Glycosylation 24-27; DEX0249_249 Asn_Glycosylation 19-22;DEX0249_250 Ck2_Phospho_Site 153-156; Myristyl 110-115;124-129;125-130;129-134;140-145; Pkc_Phospho_Site 3-5;20-22; DEX0249_251Asn_Glycosylation 40-43; Myristyl 22-27; DEX0249_252 Ck2_Phospho_Site17-20;21-24;152-155; Myristyl 22-27;28- 33;100-105;115-120Pkc_Phospho_Site 17-19;152-154;156-158; DEX0249_253 Amidation 26-29;Asn_Glycosylation 51-54; Ck2_Phospho_Site 45-48; Myristyl 50-55;DEX0249_255 Asn_Glycosylation 6-9;31-34; Myristyl 58-63;Pkc_Phospho_Site 38-40; DEX0249_256 Asn_Glycosylation 6-9; DEX0249_257Ck2_Phospho_Site 2-5;9-12; DEX0249_259 Ck2_Phospho_Site 4-7; DEX0249_260Asn_Glycosylation 208-211;294-297;601-604; Camp_Phospho_Site154-157;669-672; Ck2_Phospho_Site 247-250;338-341;352-355;513-516;519-522;527-530;551-554;556-559;606-609;637-640;683-686;739-742;747-750;756-759;795-798;817-820;871-874;885-888;898-901;904-907;912-915; Leucine_Zipper284-305;408-429;415-436;422-443;429-450;436-457;443-464;450-471;555-576;562-583;569-590;576-597;969- 990; Myristyl189-194;386-391;736-741;942-947; Pkc_Phospho_Site38-40;210-212;270-272;371-373;519-521;561-563;739-741;792-794;898-900;912-914; DEX0249_261 Asn_Glycosylation 9-12;Ck2_Phospho_Site 11-14; Pkc_Phospho_Site 34-36; DEX0249_263Ck2_Phospho_Site 62-65; Myristyl 22-27; DEX0249_264 Pkc_Phospho_Site8-10; DEX0249_265 Amidation 381-384;452-455; Asn_Glycosylation219-222;427- 430; Ck2_Phospho_Site 58-61;95-98;111-114;378-381;415-418;441-444;465-468; Dead_Atp_Helicase 40-48; Myristyl 26-31;70-75;100-105;146-151;191-196;448-453; Pkc_Phospho_Site95-97;230-232;452-454;456-458; DEX0249_266 Pkc_Phospho_Site 11-13;18-20;DEX0249_268 Amidation 26-29; Camp_Phospho_Site 28-31; Pkc_Phospho_Site14-16;40-42; Rgd 3-5; DEX0249_269 Pkc_Phospho_Site 13-15; DEX0249_271Asn_Glycosylation 39-42;45-48; Camp_Phospho_Site 9-12; DEX0249_272Ck2_Phospho_Site 11-14; DEX0249_273 Ck2_Phospho_Site 6-9; DEX0249_274Ck2_Phospho_Site 24-27; Myristyl 60-65; DEX0249_275 Amidation 201-204;Asn_Glycosylation 12-15; Camp_Phospho_Site 128-131; 193-196;Ck2_Phospho_Site 208- 211;346-349;370-373; Leucine_Zipper413-434;420-441; Myristyl 221-226;237-242; Pkc_Phospho_Site13-15;126-128;127-129;143- 145;176-178;186-188;214-216;355-357;DEX0249_277 Asn_Glycosylation 98-101;149-152; Myristyl 147-152;Pkc_Phospho_Site 151-153;166-168; Tyr_Phospho_Site25-33;26-33;72-79;75-82; DEX0249_278 Asn_Glycosylation 141-144;Ck2_Phospho_Site 21-24; Tyr_Phospho_Site 68-76;69-76;115-122;118-125;DEX0249_279 Pkc_Phospho_Site 13-15; DEX0249_281 Ck2_Phospho_Site49-52;80-83; DEX0249_282 Amidation 77-80;217-220;389-392;831-834;Asn_Glycosylation 83-86;663-666; Camp_Phospho_Site 277-280;596-599;Ck2_Phospho_Site 16-19;29-32;65-68;204-207;213-216;244-247;271-274;576-579;610-613;611-614;615-618;621-624;705-708;775-778;931-934; Glycosaminoglycan 22-25;86-89;88-91; Myristyl73-78;87-92;89-94;91-96;94-99;148-153;214-219;268-273;284-289;294-299;297-302;347-352;380-385;429-434;445-450;451-456;454-459;455-460;464-469;469-474;500-505;517-522;530-535;587-592;681-686;734-739;739-744;802-807;853-858;870-875;922-927;971-976; Pkc_Phospho_Site 77-79;166-168;217-219;264-266;327-329;440-442;472-474;534-536;588-590;699-701;716-718;926-928;940-942;956-958;972-974;

Example 6

[0514] Method of Determining Alterations in a Gene Corresponding to aPolynucleotide

[0515] RNA is isolated from individual patients or from a family ofindividuals that have a phenotype of interest. cDNA is then generatedfrom these RNA samples using protocols known in the art. See, Sambrook(2001), supra. The cDNA is then used as a template for PCR, employingprimers surrounding regions of interest in SEQ ID NO: 1 through 159.Suggested PCR conditions consist of 35 cycles at 95° C. for 30 seconds;60-120 seconds at 52-58° C.; and 60-120 seconds at 70° C., using buffersolutions described in Sidransky et al., Science 252(5006): 706-9(1991). See also Sidransky et al., Science 278(5340): 1054-9 (1997).

[0516] PCR products are then sequenced using primers labeled at their 5′end with T4 polynucleotide kinase, employing SequiTherm Polymerase.(Epicentre Technologies). The intron-exon borders of selected exons isalso determined and genomic PCR products analyzed to confirm theresults. PCR products harboring suspected mutations are then cloned andsequenced to validate the results of the direct sequencing. PCR productsis cloned into T-tailed vectors as described in Holton et al., NucleicAcids Res., 19: 1156 (1991) and sequenced with T7 polymerase (UnitedStates Biochemical). Affected individuals are identified by mutationsnot present in unaffected individuals.

[0517] Genomic rearrangements may also be determined. Genomic clones arenick-translated with digoxigenin deoxyuridine 5′ triphosphate(Boehringer Manheim), and FISH is performed as described in Johnson etal., Methods Cell Biol. 35: 73-99 (1991). Hybridization with the labeledprobe is carried out using a vast excess of human cot-1 DNA for specifichybridization to the corresponding genomic locus.

[0518] Chromosomes are counterstained with 4,6-diamino-2-phenylidole andpropidium iodide, producing a combination of C- and R-bands. Alignedimages for precise mapping are obtained using a triple-band filter set(Chroma Technology, Brattleboro, Vt.) in combination with a cooledcharge-coupled device camera (Photometrics, Tucson, Ariz.) and variableexcitation wavelength filters. Id. Image collection, analysis andchromosomal fractional length measurements are performed using the ISeeGraphical Program System. (Inovision Corporation, Durham, N.C.)Chromosome alterations of the genomic region hybridized by the probe areidentified as insertions, deletions, and translocations. Thesealterations are used as a diagnostic marker for an associated disease.

Example 7

[0519] Method of Detecting Abnormal Levels of a Polypeptide in aBiological Sample

[0520] Antibody-sandwich ELISAs are used to detect polypeptides in asample, preferably a biological sample. Wells of a microtiter plate arecoated with specific antibodies, at a final concentration of 0.2 to 10μg/ml. The antibodies are either monoclonal or polyclonal and areproduced by the method described above. The wells are blocked so thatnon-specific binding of the polypeptide to the well is reduced. Thecoated wells are then incubated for >2 hours at RT with a samplecontaining the polypeptide. Preferably, serial dilutions of the sampleshould be used to validate results. The plates are then washed threetimes with deionized or distilled water to remove unbound polypeptide.Next, 50 μl of specific antibody-alkaline phosphatase conjugate, at aconcentration of 25-400 ng, is added and incubated for 2 hours at roomtemperature. The plates are again washed three times with deionized ordistilled water to remove unbound conjugate. 75 μl of4-methylumbelliferyl phosphate (MUP) or p-nitrophenyl phosphate (NPP)substrate solution are added to each well and incubated 1 hour at roomtemperature.

[0521] The reaction is measured by a microtiter plate reader. A standardcurve is prepared, using serial dilutions of a control sample, andpolypeptide concentrations are plotted on the X-axis (log scale) andfluorescence or absorbance on the Y-axis (linear scale). Theconcentration of the polypeptide in the sample is calculated using thestandard curve.

Example 8

[0522] Formulating a Polypeptide

[0523] The secreted polypeptide composition will be formulated and dosedin a fashion consistent with good medical practice, taking into accountthe clinical condition of the individual patient (especially the sideeffects of treatment with the secreted polypeptide alone), the site ofdelivery, the method of administration, the scheduling ofadministration, and other factors known to practitioners. The “effectiveamount” for purposes herein is thus determined by such considerations.

[0524] As a general proposition, the total pharmaceutically effectiveamount of secreted polypeptide administered parenterally per dose willbe in the range of about 1, μg/kg/day to 10 mg/kg/day of patient bodyweight, although, as noted above, this will be subject to therapeuticdiscretion. More preferably, this dose is at least 0.01 mg/kg/day, andmost preferably for humans between about 0.01 and 1 mg/kg/day for thehormone. If given continuously, the secreted polypeptide is typicallyadministered at a dose rate of about 1 μg/kg/hour to about 50mg/kg/hour, either by 1-4 injections per day or by continuoussubcutaneous infusions, for example, using a mini-pump. An intravenousbag solution may also be employed. The length of treatment needed toobserve changes and the interval following treatment for responses tooccur appears to vary depending on the desired effect.

[0525] Pharmaceutical compositions containing the secreted protein ofthe invention are administered orally, rectally, parenterally,intracistemally, intravaginally, intraperitoneally, topically (as bypowders, ointments, gels, drops or transdermal patch), bucally, or as anoral or nasal spray. “Pharmaceutically acceptable carrier” refers to anon-toxic solid, semisolid or liquid filler, diluent, encapsulatingmaterial or formulation auxiliary of any type. The term “parenteral” asused herein refers to modes of administration which include intravenous,intramuscular, intraperitoneal, intrastemal, subcutaneous andintraarticular injection and infusion.

[0526] The secreted polypeptide is also suitably administered bysustained-release systems. Suitable examples of sustained-releasecompositions include semipermeable polymer matrices in the form ofshaped articles, e.g., films, or microcapsules. Sustained-releasematrices include polylactides (U.S. Pat. No. 3,773,919, EP 58,481),copolymers of L-glutamic acid and gamma-ethyl-L-glutamate (Sidman, U. etal., Biopolymers 22: 547-556 (1983)), poly (2-hydroxyethyl methacrylate)(R. Langer et al., J. Biomed. Mater. Res. 15: 167-277 (1981), and R.Langer, Chem. Tech. 12: 98-105 (1982)), ethylene vinyl acetate (R.Langer et al.) or poly-D-(-)-3-hydroxybutyric acid (EP 133,988).Sustained-release compositions also include liposomally entrappedpolypeptides. Liposomes containing the secreted polypcptidc are preparedby methods known per se: DE Epstein et al., Proc. Natl. Acad. Sci. USA82: 3688-3692 (1985); Hwang et al., Proc. Natl. Acad. Sci. USA 77:4030-4034 (1980); EP 52,322; EP 36,676; EP 88,046; EP 143,949; EP142,641; Japanese Pat. Appl. 83-118008; U.S. Pat. Nos. 4,485,045 and4,544,545; and EP 102,324. Ordinarily, the liposomes are of the small(about 200-800 Angstroms) unilamellar type in which the lipid content isgreater than about 30 mol. percent cholesterol, the selected proportionbeing adjusted for the optimal secreted polypeptide therapy.

[0527] For parenteral administration, in one embodiment, the secretedpolypeptide is formulated generally by mixing it at the desired degreeof purity, in a unit dosage injectable form (solution, suspension, oremulsion), with a pharmaceutically acceptable carrier, I.e., one that isnon-toxic to recipients at the dosages and concentrations employed andis compatible with other ingredients of the formulation.

[0528] For example, the formulation preferably does not includeoxidizing agents and other compounds that are known to be deleterious topolypeptides. Generally, the formulations are prepared by contacting thepolypeptide uniformly and intimately with liquid carriers or finelydivided solid carriers or both. Then, if necessary, the product isshaped into the desired formulation. Preferably the carrier is aparenteral carrier, more preferably a solution that is isotonic with theblood of the recipient. Examples of such carrier vehicles include water,saline, Ringer's solution, and dextrose solution. Non-aqueous vehiclessuch as fixed oils and ethyl oleate are also useful herein, as well asliposomes.

[0529] The carrier suitably contains minor amounts of additives such assubstances that enhance isotonicity and chemical stability. Suchmaterials are non-toxic to recipients at the dosages and concentrationsemployed, and include buffers such as phosphate, citrate, succinate,acetic acid, and other organic acids or their salts; antioxidants suchas ascorbic acid; low molecular weight (less than about ten residues)polypeptides, e.g., polyarginine or tripeptides; proteins, such as serumalbumin, gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids, such as glycine, glutamic acid,aspartic acid, or arginine; monosaccharides, disaccharides, and othercarbohydrates including cellulose or its derivatives, glucose, manose,or dextrins; chelating agents such as EDTA; sugar alcohols such asmannitol or sorbitol; counterions such as sodium; and/or nonionicsurfactants such as polysorbates, poloxamers, or PEG.

[0530] The secreted polypeptide is typically formulated in such vehiclesat a concentration of about 0.1 mg/ml to 100 mg/ml, preferably 1-10mg/ml, at a pH of about 3 to 8. It will be understood that the use ofcertain of the foregoing excipients, carriers, or stabilizers willresult in the formation of polypeptide salts.

[0531] Any polypeptide to be used for therapeutic administration can besterile. Sterility is readily accomplished by filtration through sterilefiltration membranes (e.g., 0.2 micron membranes). Therapeuticpolypeptide compositions generally are placed into a container having asterile access port, for example, an intravenous solution bag or vialhaving a stopper pierceable by a hypodermic injection needle.

[0532] Polypeptides ordinarily will be stored in unit or multi-dosecontainers, for example, sealed ampules or vials, as an aqueous solutionor as a lyophilized formulation for reconstitution. As an example of alyophilized formulation, 10-ml vials are filled with 5 ml ofsterile-filtered 1% (w/v) aqueous polypeptide solution, and theresulting mixture is lyophilized. The infusion solution is prepared byreconstituting the lyophilized polypeptide using bacteriostaticWater-for-Injection.

[0533] The invention also provides a pharmaceutical pack or kitcomprising one or more containers filled with one or more of theingredients of the pharmaceutical compositions of the invention.Associated with such container(s) can be a notice in the form prescribedby a governmental agency regulating the manufacture, use or sale ofpharmaceuticals or biological products, which notice reflects approvalby the agency of manufacture, use or sale for human administration. Inaddition, the polypeptides of the present invention may be employed inconjunction with other therapeutic compounds.

Example 9

[0534] Method of Treating Decreased Levels of the Polypeptide

[0535] It will be appreciated that conditions caused by a decrease inthe standard or normal expression level of a secreted protein in anindividual can be treated by administering the polypeptide of thepresent invention, preferably in the secreted form. Thus, the inventionalso provides a method of treatment of an individual in need of anincreased level of the polypeptide comprising administering to such anindividual a pharmaceutical composition comprising an amount of thepolypeptide to increase the activity level of the polypeptide in such anindividual.

[0536] For example, a patient with decreased levels of a polypeptidereceives a daily dose 0.1-100 μg/kg of the polypeptide for sixconsecutive days. Preferably, the polypeptide is in the secreted form.The exact details of the dosing scheme, based on administration andformulation, are provided above.

Example 10

[0537] Method of Treating Increased Levels of the Polypeptide

[0538] Antisense technology is used to inhibit production of apolypeptide of the present invention. This technology is one example ofa method of decreasing levels of a polypeptide, preferably a secretedform, due to a variety of etiologies, such as cancer.

[0539] For example, a patient diagnosed with abnormally increased levelsof a polypeptide is administered intravenously antisense polynucleotidesat 0.5, 1.0, 1.5, 2.0 and 3.0 mg/kg day for 21 days. This treatment isrepeated after a 7-day rest period if the treatment was well tolerated.The formulation of the antisense polynucleotide is provided above.

Example 11

[0540] Method of Treatment Using Gene Therapy

[0541] One method of gene therapy transplants fibroblasts, which arecapable of expressing a polypeptide, onto a patient. Generally,fibroblasts are obtained from a subject by skin biopsy. The resultingtissue is placed in tissue-culture medium and separated into smallpieces. Small chunks of the tissue are placed on a wet surface of atissue culture flask, approximately ten pieces are placed in each flask.The flask is turned upside down, closed tight and left at roomtemperature over night. After 24 hours at room temperature, the flask isinverted and the chunks of tissue remain fixed to the bottom of theflask and fresh media (e.g., Ham's F12 media, with 10% FBS, penicillinand streptomycin) is added. The flasks are then incubated at 37° C. forapproximately one week.

[0542] At this time, fresh media is added and subsequently changed everyseveral days. After an additional two weeks in culture, a monolayer offibroblasts emerge. The monolayer is trypsinized and scaled into largerflasks. pMV-7 (Kirschmeier, P. T. et al., DNA, 7: 219-25 (1988)),flanked by the long terminal repeats of the Moloney murine sarcomavirus, is digested with EcoRI and HindIII and subsequently treated withcalf intestinal phosphatase. The linear vector is fractionated onagarose gel and purified, using glass beads.

[0543] The cDNA encoding a polypeptide of the present invention can beamplified using PCR primcrs which correspond to the 5′ and 3′endsequences respectively as set forth in Example 1. Preferably, the5′primer contains an EcoRI site and the 3′primer includes a HindIIIsite. Equal quantities of the Moloney murine sarcoma virus linearbackbone and the amplified EcoRi and HindIII fragment are addedtogether, in the presence of T4 DNA ligase. The resulting mixture ismaintained under conditions appropriate for ligation of the twofragments. The ligation mixture is then used to transform bacteria HB101, which are then plated onto agar containing kanamycin for thepurpose of confirming that the vector has the gene of interest properlyinserted.

[0544] The amphotropic pA317 or GP+am12 packaging cells are grown intissue culture to confluent density in Dulbecco's Modified Eagles Medium(DMEM) with 10% calf serum (CS), penicillin and streptomycin. The MSVvector containing the gene is then added to the media and the packagingcells transduced with the vector. The packaging cells now produceinfectious viral particles containing the gene (the packaging cells arenow referred to as producer cells).

[0545] Fresh media is added to the transduced producer cells, andsubsequently, the media is harvested from a 10 cm plate of confluentproducer cells. The spent media, containing the infectious viralparticles, is filtered through a millipore filter to remove detachedproducer cells and this media is then used to infect fibroblast cells.Media is removed from a sub-confluent plate of fibroblasts and quicklyreplaced with the media from the producer cells. This media is removedand replaced with fresh media.

[0546] If the titer of virus is high, then virtually all fibroblastswill be infected and no selection is required. If the titer is very low,then it is necessary to use a retroviral vector that has a selectablemarker, such as neo or his. Once the fibroblasts have been efficientlyinfected, the fibroblasts are analyzed to determine whether protein isproduced.

[0547] The engineered fibroblasts are then transplanted onto the host,either alone or after having been grown to confluence on cytodex 3microcarrier beads.

Example 12

[0548] Method of Treatment Using Gene Therapy-In Vivo

[0549] Another aspect of the present invention is using in vivo genetherapy methods to treat disorders, diseases and conditions. The genetherapy method relates to the introduction of naked nucleic acid (DNA,RNA, and antisense DNA or RNA) sequences into an animal to increase ordecrease the expression of the polypeptide.

[0550] The polynucleotide of the present invention may be operativelylinked to a promoter or any other genetic elements necessary for theexpression of the polypeptide by the target tissue. Such gene therapyand delivery techniques and methods are known in the art, see, forexample, WO 90/11092, WO 98/11779; U.S. Pat. Nos. 5,693,622; 5,705,151;5,580,859; Tabata H. et al. (1997) Cardiovasc. Res. 35 (3): 470-479,Chao J et al. (1997) Pharmacol. Res. 35 (6): 517-522, Wolff J. A. (1997)Neuromuscul. Disord. 7 (5): 314-318, Schwartz B. et al. (1996) GeneTher. 3 (5): 405-411, Tsurumi Y. et al. (1996) Circulation 94 (12):3281-3290 (incorporated herein by reference).

[0551] The polynucleotide constructs may be delivered by any method thatdelivers injectable materials to the cells of an animal, such as,injection into the interstitial space of tissues (heart, muscle, skin,lung, liver, intestine and the like). The polynucleotide constructs canbe delivered in a pharmaceutically acceptable liquid or aqueous carrier.

[0552] The term “naked” polynucleotide, DNA or RNA, refers to sequencesthat are free from any delivery vehicle that acts to assist, promote, orfacilitate entry into the cell, including viral sequences, viralparticles, liposome formulations, lipofectin or precipitating agents andthe like. However, the polynucleotides of the present invention may alsobe delivered in liposome formulations (such as those taught in FelgnerP. L. et al. (1995) Ann. NY Acad. Sci. 772: 126-139 and Abdallah B. etal. (1995) Biol. Cell 85 (1): 1-7) which can be prepared by methods wellknown to those skilled in the art.

[0553] The polynucleotide vector constructs used in the gene therapymethod are preferably constructs that will not integrate into the hostgenome nor will they contain sequences that allow for replication. Anystrong promoter known to those skilled in the art can be used fordriving the expression of DNA. Unlike other gene therapies techniques,one major advantage of introducing naked nucleic acid sequences intotarget cells is the transitory nature of the polynucleotide synthesis inthe cells. Studies have shown that non-replicating DNA sequences can beintroduced into cells to provide production of the desired polypeptidefor periods of up to six months.

[0554] The polynucleotide construct can be delivered to the interstitialspace of tissues within the an animal, including of muscle, skin, brain,lung, liver, spleen, bone marrow, thymus, heart, lymph, blood, bone,cartilage, pancreas, kidney, gall bladder, stomach, intestine, testis,ovary, uterus, rectum, nervous system, eye, gland, and connectivetissue. Interstitial space of the tissues comprises the intercellularfluid, mucopolysaccharide matrix among the reticular fibers of organtissues, elastic fibers in the walls of vessels or chambers, collagenfibers of fibrous tissues, or that same matrix within connective tissueensheathing muscle cells or in the lacunae of bone. It is similarly thespace occupied by the plasma of the circulation and the lymph fluid ofthe lymphatic channels. Delivery to the interstitial space of muscletissue is preferred for the reasons discussed below. They may beconveniently delivered by injection into the tissues comprising thesecells. They are preferably delivered to and expressed in persistent,non-dividing cells which are differentiated, although delivery andexpression may be achieved in non-differentiated or less completelydifferentiated cells, such as, for example, stem cells of blood or skinfibroblasts. In vivo muscle cells are particularly competent in theirability to take up and express polynucleotides.

[0555] For the naked polynucleotide injection, an effective dosageamount of DNA or RNA will be in the range of from about 0.05 μg/kg bodyweight to about 50 mg/kg body weight. Preferably the dosage will be fromabout 0.005 mg/kg to about 20 mg/kg and more preferably from about 0.05mg/kg to about 5 mg/kg. Of course, as the artisan of ordinary skill willappreciate, this dosage will vary according to the tissue site ofinjection. The appropriate and effective dosage of nucleic acid sequencecan readily be determined by those of ordinary skill in the art and maydepend on the condition being treated and the route of administration.The preferred route of administration is by the parenteral route ofinjection into the interstitial space of tissues. However, otherparenteral routes may also be used, such as, inhalation of an aerosolformulation particularly for delivery to lungs or bronchial tissues,throat or mucous membranes of the nose. In addition, nakedpolynucleotide constructs can be delivered to arteries duringangioplasty by the catheter used in the procedure.

[0556] The dose response effects of injected polynucleotide in muscle invivo is determined as follows. Suitable template DNA for production ofmRNA coding for polypeptide of the present invention is prepared inaccordance with a standard recombinant DNA methodology. The templateDNA, which may be either circular or linear, is either used as naked DNAor complexed with liposomes. The quadriceps muscles of mice are theninjected with various amounts of the template DNA.

[0557] Five to six week old female and male Balb/C mice are anesthetizedby intraperitoneal injection with 0.3 ml of 2.5% Avertin. A 1.5 cmincision is made on the anterior thigh, and the quadriceps muscle isdirectly visualized. The template DNA is injected in 0.1 ml of carrierin a 1 cc syringe through a 27 gauge needle over one minute,approximately 0.5 cm from the distal insertion site of the muscle intothe knee and about 0.2 cm deep. A suture is placed over the injectionsite for future localization, and the skin is closed with stainlesssteel clips.

[0558] After an appropriate incubation time (e.g., 7 days) muscleextracts are prepared by excising the entire quadriceps. Every fifth 15um cross-section of the individual quadriceps muscles is histochemicallystained for protein expression. A time course for protein expression maybe done in a similar fashion except that quadriceps from different miceare harvested at different times. Persistence of DNA in muscle followinginjection may be determined by Southern blot analysis after preparingtotal cellular DNA and HIRT supernatants from injected and control mice.

[0559] The results of the above experimentation in mice can be use toextrapolate proper dosages and other treatment parameters in humans andother animals using naked DNA.

Example 13

[0560] Transgenic Animals

[0561] The polypeptides of the invention can also be expressed intransgenic animals. Animals of any species, including, but not limitedto, mice, rats, rabbits, hamsters, guinea pigs, pigs, micro-pigs, goats,sheep, cows and non-human primates, e.g., baboons, monkeys, andchimpanzees may be used to generate transgenic animals. In a specificembodiment, techniques described herein or otherwise known in the art,are used to express polypeptides of the invention in humans, as part ofa gene therapy protocol.

[0562] Any technique known in the art may be used to introduce thetransgene (i.e., polynucleotides of the invention) into animals toproduce the founder lines of transgenic animals. Such techniquesinclude, but are not limited to, pronuclear microinjection (Paterson etal., Appl. Microbiol. Biotechnol. 40: 691-698 (1994); Carver et al.,Biotechnology (NY) 11: 1263-1270 (1993); Wright et al., Biotechnology(NY) 9: 830-834 (1991); and Hoppe et al., U.S. Pat. No. 4,873,191(1989)); retrovirus mediated gene transfer into germ lines (Van derPutten et al., Proc. Natl. Acad. Sci., USA 82: 6148-6152 (1985)),blastocysts or embryos; gene targeting in embryonic stem cells (Thompsonet al., Cell 56: 313-321 (1989)); electroporation of cells or embryos(Lo, 1983, Mol Cell. Biol. 3: 1803-1814 (1983)); introduction of thepolynucleotides of the invention using a gene gun (see, e.g., Ulmer etal., Science 259: 1745 (1993); introducing nucleic acid constructs intocmbryonic pleuripotent stem cells and transferring the stem cells backinto the blastocyst; and sperm mediated gene transfer (Lavitrano et al.,Cell 57: 717-723 (1989); etc. For a review of such techniques, seeGordon,“Transgenic Animals,” Intl. Rev. Cytol. 115: 171-229 (1989),which is incorporated by reference herein in its entirety.

[0563] Any technique known in the art may be used to produce transgenicclones containing polynucleotides of the invention, for example, nucleartransfer into enucleated oocytes of nuclei from cultured embryonic,fetal, or adult cells induced to quiescence (Campell et al., Nature 380:64-66 (1996); Wilmut et al., Nature 385: 810813 (1997)).

[0564] The present invention provides for transgenic animals that carrythe transgene in all their cells, as well as animals which carry thetransgene in some, but not all their cells, I.e., mosaic animals orchimeric. The transgene may be integrated as a single transgene or asmultiple copies such as in concatamers, e.g., head-to-head tandems orhead-to-tail tandems. The transgene may also be selectively introducedinto and activated in a particular cell type by following, for example,the teaching of Lasko et al. (Lasko et al., Proc. Natl. Acad. Sci. USA89: 6232-6236 (1992)). The regulatory sequences required for such acell-type specific activation will depend upon the particular cell typeof interest, and will be apparent to those of skill in the art. When itis desired that the polynucleotide transgene be integrated into thechromosomal site of the endogenous gene, gene targeting is preferred.Briefly, when such a technique is to be utilized, vectors containingsome nucleotide sequences homologous to the endogenous gene are designedfor the purpose of integrating, via homologous recombination withchromosomal sequences, into and disrupting the function of thenucleotide sequence of the endogenous gene. The transgene may also beselectively introduced into a particular cell type, thus inactivatingthe endogenous gene in only that cell type, by following, for example,the teaching of Gu et al. (Gu et al., Science 265: 103-106 (1994)). Theregulatory sequences required for such a cell-type specific inactivationwill depend upon the particular cell type of interest, and will beapparent to those of skill in the art.

[0565] Once transgenic animals have been generated, the expression ofthe recombinant gene may be assayed utilizing standard techniques.Initial screening may be accomplished by Southern blot analysis or PCRtechniques to analyze animal tissues to verify that integration of thetransgene has taken place. The level of mRNA expression of the transgenein the tissues of the transgenic animals may also be assessed usingtechniques which include, but are not limited to, Northern blot analysisof tissue samples obtained from the animal, in situ hybridizationanalysis, and reverse transcriptase-PCR (rt-PCR). Samples of transgenicgene-expressing tissue may also be evaluated immunocytochemically orimmunohistochemically using antibodies specific for the transgeneproduct.

[0566] Once the founder animals are produced, they may be bred, inbred,outbred, or crossbred to produce colonies of the particular animal.Examples of such breeding strategies include, but are not limited to:outbreeding of founder animals with more than one integration site inorder to establish separate lines; inbreeding of separate lines in orderto produce compound transgenics that express the transgene at higherlevels because of the effects of additive expression of each transgene;crossing of heterozygous transgenic animals to produce animalshomozygous for a given integration site in order to both augmentexpression and eliminate the need for screening of animals by DNAanalysis; crossing of separate homozygous lines to produce compoundheterozygous or homozygous lines; and breeding to place the transgene ona distinct background that is appropriate for an experimental model ofinterest.

[0567] Transgenic animals of the invention have uses which include, butare not limited to, animal model systems useful in elaborating thebiological function of polypeptides of the present invention, studyingconditions and/or disorders associated with aberrant expression, and inscreening for compounds effective in ameliorating such conditions and/ordisorders.

Example 14

[0568] Knock-Out Animals

[0569] Endogenous gene expression can also be reduced by inactivating or“knocking out” the gene and/or its promoter using targeted homologousrecombination. (E.g., see Smithies et al., Nature 317: 230-234 (1985);Thomas & Capecchi, Cell 51: 503512 (1987); Thompson et al., Cell 5:313-321 (1989); each of which is incorporated by reference herein in itsentirety). For example, a mutant, non-functional polynucleotide of theinvention (or a completely unrelated DNA sequence) flanked by DNAhomologous to the endogenous polynucleotide sequence (either the codingregions or regulatory regions of the gene) can be used, with or withouta selectable marker and/or a negative selectable marker, to transfectcells that express polypeptides of the invention in vivo. In anotherembodiment, techniques known in the art are used to generate knockoutsin cells that contain, but do not express the gene of interest.Insertion of the DNA construct, via targeted homologous recombination,results in inactivation of the targeted gene. Such approaches areparticularly suited in research and agricultural fields wheremodifications to embryonic stem cells can be used to generate animaloffspring with an inactive targeted gene (e.g., see Thomas & Capecchi1987 and Thompson 1989, supra). However this approach can be routinelyadapted for use in humans provided the recombinant DNA constructs aredirectly administered or targeted to the required site in vivo usingappropriate viral vectors that will be apparent to those of skill in theart.

[0570] In further embodiments of the invention, cells that aregenetically engineered to express the polypeptides of the invention, oralternatively, that are genetically engineered not to express thepolypeptides of the invention (e.g., knockouts) are administered to apatient in vivo. Such cells may be obtained from the patient (I.e.,animal, including human) or an MHC compatible donor and can include, butare not limited to fibroblasts, bone marrow cells, blood cells (e.g.lymphocytes), adipocytes, muscle cells, endothelial cells etc. The cellsare genetically engineered in vitro using recombinant DNA techniques tointroduce the coding sequence of polypeptides of the invention into thecells, or alternatively, to disrupt the coding sequence and/orendogenous regulatory sequence associated with the polypeptides of theinvention, e.g., by transduction (using viral vectors, and preferablyvectors that integrate the transgene into the cell genome) ortransfection procedures, including, but not limited to, the use ofplasmids, cosmids, YACs, naked DNA, electroporation, liposomes, etc.

[0571] The coding sequence of the polypeptides of the invention can beplaced under the control of a strong constitutive or inducible promoteror promoter/enhancer to achieve expression, and preferably secretion, ofthe polypeptides of the invention. The engineered cells which expressand preferably secrete the polypeptides of the invention can beintroduced into the patient systemically, e.g., in the circulation, orintraperitoneally.

[0572] Alternatively, the cells can be incorporated into a matrix andimplanted in the body, e.g., genetically engineered fibroblasts can beimplanted as part of a skin graft; genetically engineered endothelialcells can be implanted as part of a lymphatic or vascular graft. (See,for example, Anderson et al. U.S. Pat. No. 5,399,349; and Mulligan &Wilson, U.S. Pat. No. 5,460,959 cach of which is incorporated byreference herein in its entirety).

[0573] When the cells to be administered are non-autologous or non-MHCcompatible cells, they can be administered using well known techniqueswhich prevent the development of a host immune response against theintroduced cells. For example, the cells may be introduced in anencapsulated form which, while allowing for an exchange of componentswith the immediate extracellular environment, does not allow theintroduced cells to be recognized by the host immune system.

[0574] Transgenic and “knock-out” animals of the invention have useswhich include, but are not limited to, animal model systems useful inelaborating the biological function of polypeptides of the presentinvention, studying conditions and/or disorders associated with aberrantexpression, and in screening for compounds effective in amelioratingsuch conditions and/or disorders.

[0575] All patents, patent publications, and other published referencesmentioned herein are hereby incorporated by reference in theirentireties as if each had been individually and specificallyincorporated by reference herein. While preferred illustrativeembodiments of the present invention are described, one skilled in theart will appreciate that the present invention can be practiced by otherthan the described embodiments, which are presented for purposes ofillustration only and not by way of limitation. The present invention islimited only by the claims that follow.

0 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 282 <210> SEQ ID NO 1<211> LENGTH: 207 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 1 ggaggagacg tgcaatagag atacccaaag aaatacatga ataattcaggagcagatttg 60 catttatcca ctggtactat ttagttgtat ttattagaca gcttcctgccctctccaaaa 120 agcttactga gctagtaact atttacaggg ttagccaaag aacacaaaaaagtgatctct 180 attagactgt aagaatatgg tttccct 207 <210> SEQ ID NO 2 <211>LENGTH: 503 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: unsure <222> LOCATION: (492) <223> OTHER INFORMATION: a,c, g or t <400> SEQUENCE: 2 ggttgatatt atattacact tctctgaata aaatgccaccttgatttgct cattctcaat 60 cggcaggagt cttatttcac ctctgtaggt cttactaagtgtgtttagtt ttcaaaagaa 120 accagtgttt ccctagtact taacatgggt ttattacatttttttgacaa aaattcaaaa 180 ttacatatat tttgttcttc attagcaagt cacacattttaaaatggcac actcccttcc 240 tcttcgtgtt gttatttgtt tattttaagg actgtttctgggtagataag ctctgggtta 300 ttttaaaata cattttacaa tggaaatggc ctggacttgaactgaaaagg aaacattatc 360 tgtgttattt cagacacatc agtgatcagt ttagaagataggatgatttc actaagctta 420 taattcatct taaagctcac ctaaataaaa gtaagtgactaaaatgatct ttttcttcca 480 ggagaggtag gnttaattaa ttg 503 <210> SEQ ID NO3 <211> LENGTH: 603 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 3 ggttgatatt atattacact tctctgaata aaatgccacc ttgatttgctcattctcaat 60 cggcaggagt cttatttcac ctctgtaggt cttactaagt gtgtttagttttcaaaagaa 120 accagtgttt ccctagtact taacatgggt ttattacatt tttttgacaaaaattcaaaa 180 ttacatatat tttgttcttc attagcaagt cacacatttt aaaatggcacactcccttcc 240 tcttcgtgtt gttatttgtt tattttaagg actgtttctg ggtagataagctctgggtta 300 ttttaaaata cattttacaa tggaaatggc ctggacttga actgaaaaggaaacattatc 360 tgtgttattt cagacacatc agtgatcagt ttagaagata ggatgatttcactaagctta 420 taattcatct taaagctcac ctaaataaaa gtaagtgact aaaatgatctttttcttcca 480 ggagaggtag gattaattaa tggtataatg tgtggaatat ttcaggcttatctgattctt 540 ccatcttaaa tctttgagag ttttaaacac attatgtgtc cattactgtttatatcacat 600 aga 603 <210> SEQ ID NO 4 <211> LENGTH: 534 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure<222> LOCATION: (133) <223> OTHER INFORMATION: a, c, g or t <400>SEQUENCE: 4 gaaatgcagc tgacaatgca aggagcaaga ggactcgcac agtggtgcatggcagcttgc 60 tgtcattttc tgggcacaga aagtgcgatg gaagggaatg agaaggggaaaaaggaagga 120 tgacaggacg ganggaggga aagaaggaag aggaaaaaag aaaggacaggagaaagggag 180 gaaggcttct gccaaaaaat taaaatcaaa tttttgacat tctttttgtttgcctttttt 240 gaaacaaaat gacacttgcc agacaccagc ttcctggccc atgtcctggtccttggtatc 300 cagatgacag cagtgtgatc ctgctgtgag ttccttccgt gccttctgatctgagttcct 360 gaaagcagag agccactcag gaactgctgt ctctcaggcc agctggctggtgatgggctt 420 ttgaagactc tggctctctc tcctgctgga agagctcccc aggggccaccaggagccagg 480 tgaccgctct cagcctctgt gagctactgg agatcaccag accttcccacatcc 534 <210> SEQ ID NO 5 <211> LENGTH: 928 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure <222>LOCATION: (329) <223> OTHER INFORMATION: a, c, g or t <400> SEQUENCE: 5atgactctta acgagcatgc tgccttcaaa catctgttta acgaagcaca tcttgcaccg 60cccttaatcc atttaaccct gagtggacac agcacatgtt tcagagagca cagggttggg 120gcactgtgcc agacactgga gataataaag aaaaacagca ctgggcctat aattgggagt 180ctagatatac gatatggaaa tgcagctgac aatgcaagga gcaagaggac tcgcacagtg 240gtgcatggca gcttgctgtc attttctggg cacagaaagt gcgatggaag ggaatgagaa 300ggggaaaaag gaaggatgac aggacggang gagggaaaga aggaagagga aaaaagaaag 360gacaggagaa agggaggaag gcttctgcca aaaaattaaa atcaaatttt tgacattctt 420tttgtttgcc ttttttgaaa caaaatgaca cttgccagac accagcttcc tggcccatgt 480cctggtcctt ggtatccaga tgacagcagt gtgatcctgc tgtgagttcc ttccgtgcct 540tctgatctga gttcctgaaa gcagagagcc actcaggaac tgctgtctct caggccagct 600ggctggtgat gggcttttga agactctggc tctctctcct gctggaagag ctccccaggg 660gccaccagga gccaggtgac cgctctcagc ctctgtgagc tactggagat caccagacct 720tcccacatcc cgggcaggtg ccagggcctt taaggaggct ttctgctctg cagggatgtt 780ctgtgggctc cagtattctg gcgagcatca gcttattctc ggcttagtct tcttgctcta 840tagctcctgc ctctgttttg cttctttttg gtgatgcctg tttcacctta tgaagagggc 900ctgtaaaatc caagctctgc acaaaccc 928 <210> SEQ ID NO 6 <211> LENGTH: 368<212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: unsure <222> LOCATION: (305) <223> OTHER INFORMATION: a, c, gor t <400> SEQUENCE: 6 tattaagtga ctttaatgta agttacccac ttctggtgctagtttgtatg ttaatttgtt 60 tttatcctga tacatataat caggtagaat tgggaattttattcagcctc cgcgtcgggg 120 aacatagaat aactctttat tgactcaaga ctggaattttcagaaaagtt taaattttta 180 ttttttctag aattttcaga aaagtttaat agatctgagacatttttaaa tcttttaatc 240 tttctagcta tttgtgaata tgcttttctt ccttttaaataaatataata gctggtatgt 300 aaganagcta ttgatgcata tttttatttg gatatcttattgaactctta attggaataa 360 ttttcagt 368 <210> SEQ ID NO 7 <211> LENGTH:583 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: unsure <222> LOCATION: (61)..(80) <223> OTHER INFORMATION: a,c, g or t <221> NAME/KEY: unsure <222> LOCATION: (495) <223> OTHERINFORMATION: a, c, g or t <400> SEQUENCE: 7 cacagcagca tctggactggtatctgacca aaaactaggt accatggtct agccacattt 60 nnnnnnnnnn nnnnnnnnnnatagtcataa gatttggttt tctcttactg aaatgcggta 120 taattgggat ttattcaaatcttcttttat atccatccca taaagttttg taatttttcc 180 ttattggcct aacaattcctgtaaagatta tttgtaagta gttaataatt tttgtaatca 240 tgatgagttt aacacttttattttattaca tgtgctcata tgttgttgct gacactaaga 300 aacctatagg tcaaaagatgcaaactaggg ccacatgagc acggcagcac ccagccagga 360 ctctgctgca gctgccgtttgtagatggag ctctctgtct ccaaagaagc acaggcctgt 420 tgttcttctg gttgtgctacagtaaaatga acctgggttt tctgaacatg tggttgaatg 480 tcagttgcta cctancgtttcacttgtgat attgatttta ttaatcttaa agttatgtga 540 atgttaacta tttctcatatctatatctta ttcaaaatct ggg 583 <210> SEQ ID NO 8 <211> LENGTH: 118 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 8 gaaaaaaatcctgaaatatt caaatgttta ccttactaaa ggaattctta atctgctgat 60 tatgtttcttaattgtacca tgaattacaa aaacctattg gcaagatcag tcttattt 118 <210> SEQ ID NO9 <211> LENGTH: 502 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 9 tccacttttt aaaaagagtt agttatatgt agcatagtaa aaatatttcttttgaaatat 60 actattatat ccatttattt aagttctaat agactatact gcctaaaaaaatagagttga 120 tcttagtcaa atcacttaaa tcaggaacct ttaagattca gtttaatcagttttgtatat 180 aaagtattag acttactttc accatttttg cccaaaaaca aacttctgtacttcattaaa 240 taacatcaac aaaagtaaaa agctaacatc aacctaggaa aaccattttcaccgtaacag 300 ttatgtatat taactatatt atatactttt aaattagtaa aaaaaagggtaaaagacatg 360 aacaattatc aaaggaaaaa aatcctgaaa tattcaaatg tttaccttactaaaggaatt 420 cttaatctgc tgattatgtt tcttaattgt accatgaatt acaaaaacctattggcaaga 480 tcagtcttat ttaaaaaaaa aa 502 <210> SEQ ID NO 10 <211>LENGTH: 390 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: unsure <222> LOCATION: (83)..(211) <223> OTHERINFORMATION: a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (214)<223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY: unsure <222>LOCATION: (300) <223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY:unsure <222> LOCATION: (304)..(324) <223> OTHER INFORMATION: a, c, g ort <221> NAME/KEY: unsure <222> LOCATION: (368) <223> OTHER INFORMATION:a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (381) <223> OTHERINFORMATION: a, c, g or t <400> SEQUENCE: 10 cacatatatc cagtaacagggctgtgcgtg taccctctaa atctacaaaa ataaaaagtc 60 ctcaagtttg acttattgtttannnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120 nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180 nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn nacncgtatt cctttctgga gactctaggg 240 gaatctcttt tcttgccctttctggcttct agaacctgcc tacattcttt ggctagtggn 300 cccnnnnnnn nnnnnnnnnnnnnngtggct ggtcagtctt tctctgatgt tatctttctg 360 gttctgancc ttccatctccntcttcgcca 390 <210> SEQ ID NO 11 <211> LENGTH: 266 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 11 cttcagtggt aagagttctcatattaaatt gaattggtac ttaagaaagg gctttaacta 60 ggcaaaacct tgggagttttgaggtccctg tggacttgcc tggcatgcct gggagatttg 120 taggtatctt ctttctacacggattgtgtt tctgccctca aggaaaatag ttcactttga 180 ccactgtaaa tgatgtagtatttaaacaaa aggaaaagca cttcattgtc tgctctaaaa 240 ctaaaatgtt aagaaagagaggtggc 266 <210> SEQ ID NO 12 <211> LENGTH: 380 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 12 tgtcatggga agtggcggccactgccacat ctgctatgag gctcttcgcc atcgttggct 60 gctggaaatt tggatacagtaaatggtata taaggcttct ttttgcttgt gcaccagaag 120 tctttgtccc agcctccagatcagcagtca gtactcccct ctcccaacct gtaggaagta 180 cttgtgaaaa gttatctatccctggtctga gtgggaggtt cttaacctca ttgatgtttt 240 agtgtgactt gtctacatttgtgtgctccc ttcgtcatct gcagaggata tgagaaaaga 300 aacaaatgaa caaaaagtggagatagcgcc ttcctattca ttcttcattt ggtatggtta 360 tagttaagag aggtgagcca380 <210> SEQ ID NO 13 <211> LENGTH: 871 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 13 tttttgctaa gacaagatct actttgtcacccaagctgga gtgtagtggc gcgatcatgg 60 ctcattgcag cctcggtctc ccaggctcaagcaatccttc cacctcagcc tccagactag 120 ctggaactac aggcgcatat caccaagcctggctaatttt ttaattaaga cagggtctac 180 tatgtggccc aggctggtct tgaactcctggactcaagca attctcccac tctggcctcc 240 caaagtgata ggattacagg catgagccaccacgcccagc ctggctcacc tctcttaact 300 ataaccatac caaatgaaga atgaataggaaggcgctatc tccacttttt gttcatttgt 360 ttcttttctc atatcctctg cagatgacgaagggagcaca caaatgtaga caagtcacac 420 taaaacatca atgaggttaa gaacctcccactcagaccag ggatagataa ctttcacagt 480 acttctacag ttggagaggg agtactactctgatctggag gctggacaaa acttctgtgc 540 acaagcaaaa aaagcctatt accattacctgtatccaaat tccagcagca acgatggcga 600 agagcctcat agcagatgtg gcagtggccgccacttccca tgacaaaggg aggtaggcat 660 gattcatctc tagtgcatgg ggacggttgccttttgcccc aaatcaactc aagacccctt 720 tccaccatat gtctgtatgt aactctaaatgcatctctaa gacttaagaa taaaaagcca 780 ctgctacctg gcaagtggat gaggcaaaagagaagacata ccccaaagaa ctatagcact 840 ctgctccaaa ttacagaact ttctaaacgt c871 <210> SEQ ID NO 14 <211> LENGTH: 411 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 14 gaacaaacct ggaatattgt gaactattgatgacaatgct ttgagggcat ctttggaaac 60 caaaatgtaa atataactaa ttattttttcacttattgtt agcatttagt attttatgca 120 taaaaacttt tttaccaaat aaattttggaagtttaaatt ccacaaatga tactaatgaa 180 agtataaatc attttgggtt gttttttaaaaaattatgtt tcaatctgtc attattggaa 240 taaagtgtat aaactgcatg ttataaaacggctttacaca tatataactc atgaactcaa 300 gagaaataat atttttagga aagaagagtatctctagatt tttaataata attaaatttc 360 tttaaaagac tgaagataga aaagagaaattaaactatgt attgacttaa t 411 <210> SEQ ID NO 15 <211> LENGTH: 737 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:unsure <222> LOCATION: (657) <223> OTHER INFORMATION: a, c, g or t <221>NAME/KEY: unsure <222> LOCATION: (709) <223> OTHER INFORMATION: a, c, gor t <221> NAME/KEY: unsure <222> LOCATION: (716) <223> OTHERINFORMATION: a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (721)<223> OTHER INFORMATION: a, c, g or t <400> SEQUENCE: 15 atcaaacaaggcaaggagaa ctcaagaaag tctggtcact tcagtgagtg ttggtgtgtg 60 aggagaatcaagtgagaagt gcttttagaa acatccatgc aggtatgaag gagctctaac 120 atgccaggtgagtggacaag gcttggggaa ggttgacgat acctagagac atgtcagcct 180 ttgggtcagagcctgcctaa tctcatgggg aactgatggg tggaataggc actctaggct 240 ccccctggcatgggcagggt tgcagagaat tcagaaattt gtgtgtgttc acggagggga 300 ggccaagcttctgactgttg aagttagggt cagaatggag aggccagcat gtgtgtgagg 360 gactctgagatgggaggctg acaattagga caatgggctt tgcccatgga gccagaatgt 420 taccctgcgagggtgaatgg cactgggcca catatctcag tggtcagatc caggatctca 480 gcagtcactcctatcaatca gagatcggac catggcaggc acactcagca aagtccccca 540 tgatccagaagacatgtgcg aattctgcat catcttcccc tccataatcc tgaggacagt 600 gagagccaaggtaaggacct tgacacacag attcgttacc aggaggaatt ctttgcntac 660 tgaatcattctgaatatatc ggatttgtct aaatagatcc cacctgccnt cccatntact 720 nttctccagtaaagagg 737 <210> SEQ ID NO 16 <211> LENGTH: 1082 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure <222>LOCATION: (90) <223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY:unsure <222> LOCATION: (1002) <223> OTHER INFORMATION: a, c, g or t<221> NAME/KEY: unsure <222> LOCATION: (1054) <223> OTHER INFORMATION:a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (1061) <223> OTHERINFORMATION: a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (1066)<223> OTHER INFORMATION: a, c, g or t <400> SEQUENCE: 16 ttatgggtagatattatgat gacgtatata ttctacagat ggagaaacta tgacctagag 60 tgaccccaatgtcaccagct aagggtgggn ctgggtttaa accaggccag tctagcttgt 120 aacctctatgtcatattgga atagactcta gcacagtggt tataggttcc tactaaaaga 180 gtgttaaaaccatgacagct atgctttaag aatacatact tgaatgtaag tgttagccct 240 gacatccaatttgtcttcct aacgcctagt atataatatg gctcaagaaa tgtttattaa 300 atatgtgtttcatttttttt ttcttttttg agatggagta tcactctgtc acccaggctg 360 gagaactcaagaaagtctgg tcacttcagt gagtgttggt gtgtgaggag aatcaagtga 420 gaagtgcttttagaaacatc catgcaggta tgaaggagct ctaacatgcc aggtgagtgg 480 acaaggcttggggaaggttg acgataccta gagacatgtc agcctttggg tcagagcctg 540 cctaatctcatggggaactg atgggtggaa taggcactct aggctccccc tggcatgggc 600 agggttgcagagaattcaga aatttgtgtg tgttcacgga ggggaggcca agcttctgac 660 tgttgaagttagggtcagaa tggagaggcc agcatgtgtg tgagggactc tgagatggga 720 ggctgacaattaggacaatg ggctttgccc atggagccag aatgttaccc tgcgagggtg 780 aatggcactgggccacatat ctcagtggtc agatccagga tctcagcagt cactcctatc 840 aatcagagatcggaccatgg caggcacact cagcaaagtc ccccatgatc cagaagacat 900 gtgcgaattctgcatcatct tcccctccat aatcctgagg acagtgagag ccaaggtaag 960 gaccttgacacacagattcg ttaccaggag gaattctttg cntactgaat cattctgaat 1020 atatcggatttgtctaaata gatcccacct gccntcccat ntactnttct ccagtaaaga 1080 gg 1082<210> SEQ ID NO 17 <211> LENGTH: 128 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 17 gtttttgcaa ataagagcac taaaaagactaaaccattcc tcggtgcctg gaagaggctg 60 tttgcatttt agttaccctg ctgttcataacatctctaag aaaatgtagg ggccaccctg 120 ggcgcagt 128 <210> SEQ ID NO 18<211> LENGTH: 465 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 18 gaaggagaga gtgagtgagc aaatgactga aagcagccag cattgccgggaccctgccgc 60 atctcatact gggtgctgca caggcagcgt tcctcctctt gtggcgcttgggaaatagac 120 attaatcaca cacaaacaaa aacgatggca aattgtaatg agggctatgaaagggagtga 180 agggcaagat cactgagggg tgggactcag aagaatcctt tctgaggacatgacctgtca 240 gctaagacgg aaggaggact tcagttgaga gaacaagtca attaatactccgtggcctct 300 tccactcact ctcatgaatt tgccaagtcg cctggggaga atggggcctgaactctggac 360 cctgaggtct ttctccatgc actgaaagca gcccccccag ccctgggcccctccctgcga 420 cttggtgact tgactgccct attctagcat catggaaaac acatg 465<210> SEQ ID NO 19 <211> LENGTH: 539 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 19 gtaaaaagga aatttgttgt cattctgtggaatagagtga ttaagcagct tgcctaaact 60 cccccagaca gggatgcagt tggagctagaaagttaaccg aggcctggct gacaaggcct 120 cattctccat cagctggtca aaggaacaggagctctgcat cctgtcctgc tcagtagagg 180 aggaaaggag acagctcctc cgggaaatctagaggaattt gcattctctg cctgagctgg 240 cttggggcca gctttgtgga atcctggtgccagttctttt tctgcttcag gggctatttt 300 ttaaaaaaat cagtagttac attttatgtttaccaataga tttatatagc aaatgatatt 360 tgttttttat ttaaagccac aatatcaagtgtctttttaa aaatataaat aataatcctc 420 atggtatgca gatgtagcag aaaattgtgcaggtggtatg tgggcaactg gatttgggga 480 aatgctgctg catgtcatgc actctccataggtaggtttt cccctttatt tcctccctc 539 <210> SEQ ID NO 20 <211> LENGTH: 641<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 20gtaaaaagga aatttgttgt cattctgtgg aatagagtga ttaagcagct tgcctaaact 60cccccagaca gggatgcagt tggagctaga aagttaaccg aggcctggct gacaaggcct 120cattctccat cagctggtca aaggaacagg agctctgcat cctgtcctgc tcagtagagg 180aggaaaggag acagctcctc cgggaaatct agaggaattt gcattctctg cctgagctgg 240cttggggcca gctttgtgga atcctggtgc cagttctttt tctgcttcag gggctatttt 300ttaaaaaaat cagtagttac attttatgtt taccaataga tttatatagc aaatgatatt 360tgttttttat ttaaagccac aatatcaagt gtctttttaa aaatataaat aataatcctc 420atggtatgca gatgtagcag aaaattgtgc aggtggtatg tgggcaactg gatttgggga 480aatgctgctg catgtcatgc acctctccat aggtaggttt ttccccttta ttttctccct 540cttttcagca aaactctccc ctctactttc ccactgaaaa atagcatggt gagttaaact 600agttagtctg atcatggtgt gggcatggtg gttgggttag a 641 <210> SEQ ID NO 21<211> LENGTH: 406 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (379) <223> OTHERINFORMATION: a, c, g or t <400> SEQUENCE: 21 ctggtagtca ggggcaaactcaggccagaa cccaagtttc ctgactacta ttccagtgct 60 ctttccaccg tgctctggaagaatacggca caagcataag agcgtagtat tcatccctgt 120 acattcatgc agccagccttacctagagtc acagtcaatt gtggccaact tggcaagatt 180 tgaacatcac tgataagcaatctttctctc aatgctgcat ctctccagct tgttctttcc 240 ctaccatccc ccacgtatgactaaagttat agcattgact gaaatctttg ggattaaagc 300 cctgtgatct gactgagaaaaacctgttga gccattacct acaatttaca caaacaaatt 360 tcttcgattt gtcttttanggctggcccga aggcatttac atttga 406 <210> SEQ ID NO 22 <211> LENGTH: 467<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 22cagacagaac agatagatag aagaaaagaa aagggtcact tggcactagg tcttcacagg 60taaagattca gagtgtgata ggaagcacag gctcaggcac ccgggtctaa tcaatgacaa 120tctcgcttct aggccttttg gtggcatttt ctagtctacc tctaagctct agggaatcgt 180gtggctaaaa tcttccctcc tgctgagact cagagaatac catgttggcc aagatctcta 240aaacaatcaa acctggcagt attgagttac cttcctctta tcataaagtc tttcctcact 300tcctccttat tgtgaacttt cttaagaagt gagtccagga ggaagcagtg acatgaattt 360attaacttga ctcagacttc taaagacaac acaaactggg cgccccattc agagagtgac 420agggaaaccc cgtggcataa ttagttacta cgagtttcca aatagga 467 <210> SEQ ID NO23 <211> LENGTH: 1328 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (300)..(472) <223> OTHERINFORMATION: a, c, g or t <400> SEQUENCE: 23 gcacagattt agccttggtatttttttctg ggaagtataa aagacttttg tgttctgtct 60 ttttgttttc aatttctctctagaggaatt taaaaccgga tatttccatc ttaaagttct 120 tgagcaagtc tgtcaaggtgtccatatttc ttaccctgtt cctctcagca tcgaagtgct 180 atctctgtta cactcatgtttgctgttcac aatggagtac taatgaaata gcaaaattaa 240 gctaccggca tggtgctaataactgaaact aaaaatcggt ttggagcttt tctgtttggn 300 nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 360 nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 420 nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnaagaaaag 480 aaaagggtca cttggcactaggtcttcaca ggtaaagatt cagagtgtga taggaagcac 540 aggctcaggc acccgggtctaatcaatgac aatctcgctt ctaggccttt tggtggcatt 600 ttctagtcta cctctaagctctagggaatc gtgtggctaa aatcttccct cctgctgaga 660 ctcagagaat accatgttggccaagatctc taaaacaatc aaacctggca gtattgagtt 720 accttcctct tatcataaagtctttcctca cttcctcctt attgtgaact ttcttaagaa 780 gtgagtccag gaggaagcagtgacatgaat ttattaactt gactcagact tctaaagaca 840 acacaaactg ggcgccccattcagagagtg acagggaaac cccgtggcat aattagttac 900 ctacgagttt ccaaataggatttggaagga gacatacaac taggtcgccg gcgtggcaca 960 tggcttccct gaagccagcattgcctggcc aaggaagctt tgcagaacag atgagatttc 1020 agctgggact tgcagccaagtgggatttgg ccttttgggg agaagggaaa gggcattcaa 1080 aggccaggga cagagtatggtcaaaggcat ggagatgagg aagaggggac cagagcagag 1140 ggtcaggttg gaaagcgagttggggtcaat ctgcaaaggg gctgacgtgc caggtaaaaa 1200 acaggagcac cgtttagttttgtcggatca tttcaggtgg aagggcagtg ggaatgttgg 1260 agaaaacact ttttggtgtcgttacattga atctgctcat ctataagaat aaaactttat 1320 ttcataga 1328 <210> SEQID NO 24 <211> LENGTH: 550 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (352)..(371) <223>OTHER INFORMATION: a, c, g or t <400> SEQUENCE: 24 ctcatacctc ccctggtccagatcatgatt caggtctttg ttctaggatt cctggcttat 60 tatctggttt atttcaataacaaggacaat aagtcatggt tatgattttt ctgtttcatg 120 gagtgagtga acattttatctcattccagg aatttgtttt tttccaacta ttgttgcttt 180 ttgggttggt tttaaatattcctttaccaa gaaattcatt cattagtcta cattttcagc 240 tttattagca taggagtcttaataacattt tgtgtatgtt ttcatcacct agtgattgtt 300 tgctaattcc tcaccttttctttaggtcac tgttctttat acattggtta tnnnnnnnnn 360 nnnnnnnnnn ngcatttaagtttttacttt ttatgagaca aatgtatttg cgttccatag 420 atgtcagtta gaaatgttttcaacatcatg gttctctaca aactttgtga tttcagttac 480 atttccacat tgactcaacagttatttaat agtgagctct ttttttttta agacgtagtc 540 tgactctgtt 550 <210> SEQID NO 25 <211> LENGTH: 150 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 25 gattcagccc gtgatccttg actggatcct gggtgaaagcaaaagcagct ctaaaggaca 60 ctttgcagac taaatgttag ctaacaccat tgtatcagtgagaaagtgca gagtgtggtg 120 agtccattga ggctctgtag aagaaagtcc 150 <210> SEQID NO 26 <211> LENGTH: 192 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 26 aaaaagtcaa tgtcataaaa gacaaagaaa ggctgaagaagtgattcagc ccgtgatcct 60 tgactggatc ctgggtgaaa gcaaaagcag ctctaaaggacactttgcag actaaatgtt 120 agctaacacc attgtatcag tgagaaagtg cagagtgtggtgagtccatt gaggctctgt 180 agaagaaagt cc 192 <210> SEQ ID NO 27 <211>LENGTH: 747 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:27 gagctttgca gggatttagc ttttctcagg gccacctgcc ctcaggcttc ctgggccctc 60atacttcttc ttgtttatat cttatctgcc tttgggggaa tgaccttaga ggaattggtg 120tgagtaagcc atgaggttct tggtccacct ccatccagcc aagggcagct ggcagctggg 180cacttacatc cagcaaggca gaagcaaccc tggctttgaa gtcagactgc tagggtgagt 240ctgaatggcc tcggggaaag ttccctctga gccttcgttt ttttcacttg tgaaggcgat 300agtctcgcct agcttgaggg tttatcaggg ggattcagtg agaacctcat ttgaagcagc 360tgctttagtt cctaacacct aataaatgtt taaccactta ccctcctctc ccaccaccct 420ttcaactttg aacctcttcc tccatgtcat cccttcttaa ggcgctgacc ttttggccac 480aaagaatggc tctttttgtt cccatcagga ctagaattct tatctttttg ttgcttggcc 540ctggtaatca aagaaccacc aacacatttg caaggcatct ccagccttct cgttctggcc 600gcccctctct gtcttaggga gagtgctata ctggcatggt gatgagatga acgaaagggc 660agtctctggc tgttttctgc tgatgaggat gtgctgagca gcctcctgca aatgagaagc 720agggaaaaga ccaaactagc ttagctc 747 <210> SEQ ID NO 28 <211> LENGTH: 184<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 28taagctcgga attcggctcg aggcatgcca gtctttgggg catatggatg gtggatgtgg 60cttgcctttt caccctctga cattgttgat gagcagaaac ttccaatttt gatgtagtcc 120acaatattaa tatttctcat aggccaccac acccagccta tgttatcttt tagaagcttt 180attg 184 <210> SEQ ID NO 29 <211> LENGTH: 217 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure <222>LOCATION: (97)..(161) <223> OTHER INFORMATION: a, c, g or t <400>SEQUENCE: 29 ctcaattatt ctaggaatct atgctgaata tgcctctaac aatacaaattatgtattact 60 taatgttatt aattatagta ttatttaatt tgcgaannnn nnnnnnnnnnnnnnnnnnnn 120 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn ntagccatataatggagtac 180 catgcagata aatcagaaac tagagattgg ttacctg 217 <210> SEQ IDNO 30 <211> LENGTH: 543 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 30 tgaggaagcc tgaatgggaa tgaaaaacaa ataagaatcaaaagcagagg gctgtgtctt 60 ttatttttgt attcttacac cgtatgaact ttattaagaactaaaatcat gtgatggtaa 120 cccatgggca cacgctgagt acaaagttgg aaaattgtaccagcatcatt aactggggtt 180 gctttgtgtt aacattgtgt atcattcaca gatgtcaacaataagagaac acatctctct 240 atatataatt gttacaaaca ttttaaatta taaggaaaagaaaaaaaaag atgcaaaagt 300 tcaaagacta aactcacaac atcctaccga cagagaatacttaggatagc taagatctca 360 gtgttcattc aaacttctaa actcaaagac tcggatttatgcaggaagtt gaacatgtct 420 tgtgtcacca gatctgtcag tgaagctgat tatagtcttgggataaattt gagagtaatg 480 tgacagtggt ctgggaactc tgactgtggg ctctgccctctggatgcaga gactccaagg 540 cac 543 <210> SEQ ID NO 31 <211> LENGTH: 283<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 31gcttcaagct tcctgctctc ctcttttgcc atattattga gcctggaatc tgagtgggag 60aggacgacag agggtctggg cacaaggaag ccattgattg aggccattac tgcaatcaac 120ccaccaccaa taaaaagcac tggaggagag ggcttgacat agacacaaaa cataaaggaa 180gggggtgaaa ggaaggaaag agattgagga aaaaaaataa aaataaaaga tggctgggta 240agggaagaag agatagggaa gagagacaga aaaggtagaa tgc 283 <210> SEQ ID NO 32<211> LENGTH: 418 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 32 aaaaaatcct tcatgtgtct tgtatacatc tgcagaagac cagtagtgtacattttcttg 60 gctgtagcaa gggtccaaga aaggaagtag taagcagttt ttcaagtctctctctctctt 120 tttttatttt gttggcttca atgcttcctg ctctcctctt ttgccatattattgagcctg 180 gaatctgagt gggagaggac gacagagggt ctgggcacaa ggaagccattgattgaggcc 240 attactgcaa tcaacccacc accaataaaa agcactggag gagagggcttgacatagaca 300 caaaacataa aggaaggggg tgaaaggaag gaaagagatt gaggaaaaaaaataaaaata 360 aaagatggct gggtaaggga agaagagata gggaagagag acagaaaaggtagaatgc 418 <210> SEQ ID NO 33 <211> LENGTH: 172 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 33 cagactggga ctctggaaaatcctaaagca ttatagaact tggggcttgt cctttgactt 60 catggttttc aaacccagcatggtgaccca gtagtgggct gtccaatcaa ctgagctctt 120 gaaattggaa tagaataaaatagaaatatg agcatattcc catctataga aa 172 <210> SEQ ID NO 34 <211> LENGTH:128 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 34ggcctccgat tgtcccacag ttagttgttc ctcggaggca cccctcctgc tgctccttgg 60atactccagg ggccgaggag ccgagactca ctggagtgtg ggcatggcca tccagagagc 120tctgatca 128 <210> SEQ ID NO 35 <211> LENGTH: 619 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure <222>LOCATION: (128)..(416) <223> OTHER INFORMATION: a, c, g or t <400>SEQUENCE: 35 ggcctccgat tgtcccacag ttagttgttc ctcggaggca cccctcctgctgctccttgg 60 atactccagg gccgaggagc cgagactcac tggagtgtgg gcatggccatccagagagct 120 ctgatcannn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn 180 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn 240 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn 300 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn 360 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnngctt 420 tggtctttgg gggttgctga aaaagcaaaa ccaggtctgt ggggtagaaggcgccctggc 480 cacacacagg cattgccgcc tctggggtcc gcagagtctg tgtgacaacctggtcactcc 540 gatctagcag cgtatttgaa tgaatgagtg acagcttaat gaagtagccaagtaccttga 600 tttgaacgta ggagccggg 619 <210> SEQ ID NO 36 <211> LENGTH:356 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 36cgacagataa gtcagatatc gaatatagac attaaaagat ctggggcact aggctgtacc 60ctgttattgt cagtggctct ttagtcctta aacaagggtc ttgcctccta cttttttttt 120gttatggtag aaataaatgc ccacccgagt tttcatcact cactattatt ctatctttgg 180tgtgcctgta ccatgttccc ttaacaatcc tcaattatga aacatttagg cagtttataa 240acaatactgc aatgaacaac ctagtgcata ctttttttgt gtgttcttct tttattattt 300cctagaagtg agccctagaa atggagttcc tgagtcaaaa tgacacattt tatagc 356 <210>SEQ ID NO 37 <211> LENGTH: 158 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 37 aaggaattag attccacatc tcaatctaag gagcagcacaaatatgcaga gaggaaagga 60 attgattgtg gccctctttg aaaactatct caggccatccttgggccact tcaattcata 120 gctcttcctt atgcaaaata cactcacctc ttgcattt 158<210> SEQ ID NO 38 <211> LENGTH: 585 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION:(159)..(279) <223> OTHER INFORMATION: a, c, g or t <400> SEQUENCE: 38aaggaattag attccacatc tcaatctaag gagcagcaca aatatgcaga gaggaaagga 60attgattgtg gccctctttg aaaactatct caggccatcc ttgggccact tcaattcata 120gctcttcctt atgcaaaata cactcacctc ttgcatttnn nnnnnnnnnn nnnnnnnnnn 180nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnt ctcatgtgac tgcagtcaag 300gtgtcagatg agctatattc tcatctggag gcttgacagg ggaagagcct acttccaagt 360tcattaggtt ggtggtagaa ttcactccat tgtggatgta tgactgagga ccctggcttt 420ttgctagcca tcagaagagg ccagtcttgg gtcctagaga ccacctgtgg ctcccttaca 480atgtgggctt tctcaacatg gctacttact gcatgaagcc agcaaaaaga atctcccagt 540ccagtatgct aagacagagc cttgttataa cataagtcca ccctc 585 <210> SEQ ID NO 39<211> LENGTH: 295 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 39 ggggggggca gtagtttctg aagagagagc taaactgcat gagcagatgcttagccaatt 60 tctaaaaatg gaatgggagg tagaaatttc acaggtggtt gctggtttgcagcatttcca 120 catactagga tacatcatca caagatgttg tctgccagct ggtgctataactgctagtaa 180 agccacttgc ttctgaatgc atggtgatag tagtgaatcc ctaatgtcagtgcaatgctt 240 tacttatttg ctataaaatc ctttcatagt cagaagcact gttgtgttcctggca 295 <210> SEQ ID NO 40 <211> LENGTH: 302 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 40 gcgggggggg gggcagtagtttctgaagag agagctaaac tgcatgagca gatgcttagc 60 aatttctaa aaatggaatgggaggtagaa atttcacagg tggttgctgg tttgcagcat 120 ttccacatac taggatacatcatcacaaga tgttgtctgc cagctggtgc tataactgct 180 agtaaagcca cttgcttctgaatgcatggt gatagtagtg aatcccttaa tgtcagtgca 240 atgctttact tatttgctataaaatctctt tcatagtcag aagcactgtt gtgttcctgg 300 ca 302 <210> SEQ ID NO41 <211> LENGTH: 346 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 41 aagtaattaa cttgatcaaa ctcattttac agatgaggaa actgattccaccctccatgc 60 tcttcacctg cattctaaac tcttccaggg ccctccttac caggcagaggcaaattgagg 120 aagtggacac agcatttcct ttccttgttg tttgacatgc aaagcactttagactatatt 180 tagtacctaa ttgatgtggc agcaggggcc gcctgggatg ttgtggcatcatttttgctc 240 tcaatgagac acgataggga tggtttggtg gtggtttcaa aactaaagaccctccagcag 300 agcctgtcaa gtaaaacaag ggtgactgct tggttgccat accagg 346<210> SEQ ID NO 42 <211> LENGTH: 468 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 42 aagtaattaa cttgatcaaa ctcattttacagatgaggaa actgattcca ccctccatgc 60 tcttcacctg cattctaaac tcttccagggccctccttac caggcagagg caaattgagg 120 aagtggacac agcatttcct ttccttgttgtttgacatgc aaagcacttt agactatatt 180 tagtacctaa ttgatgtggc agcaggggccgcctgggatg ttgtggcatc atttttgctc 240 tcaatgagac acgataggga tggtttggtggtggtttcaa aactaaagac cctccagcag 300 agcctgtcaa gtaaaacaag gttgactgcttggttgccat accaggcaca ggttagcatg 360 aaacaaagtg tagtgtccaa ggagagggagcagggtgtct cctttgggtg agctttgcaa 420 ggggacttgg gacttggctg gaaaaggtgttttttttagt tgtatgtt 468 <210> SEQ ID NO 43 <211> LENGTH: 107 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 43 ttcaccgtgctgtgtgaatt gtggctttaa atgtattcct gtcaattcca tatattttta 60 aaatgttgcttttagagtat gtgcaagttt ggggcatttt tgagggc 107 <210> SEQ ID NO 44 <211>LENGTH: 352 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:44 gaacatgatt gagttagaaa ccagtgtggc ctgggactgg gaagctcatt aaaggaattg 60ggacttaaac tgggaagggc aagttggctc tagatccata gaaactgaag acaggggaag 120agagagatgg tattatagat ggaagaaggg gcagtgggtc atggaataaa tattggtgag 180caggggagca aaccaaaggg gtaattggga gattctgagt tttcaaggct attaaaatgc 240agttccaggc cctagggagg agagttccag actgttttct ctacactgct ataattcctt 300acactgctgg gagcagtttc tttgacatac tttgcaactg cagagggctt tt 352 <210> SEQID NO 45 <211> LENGTH: 356 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (98) <223> OTHERINFORMATION: a, c, g or t <400> SEQUENCE: 45 gaacatgatt gagttagaaaccagtgtggc ctgggactgg gaagctcatt aaaggaattg 60 ggacttaaaa ctgggaagggcaagttggct cctagatncc catagaaact gaagacaggg 120 gaagagagag atggtattatagatggaaga aggggcagtg ggtcatggaa taaatattgg 180 tgagcagggg agcaaaccaaaggggtaatt gggagattct gagttttcaa ggctattaaa 240 atgcagttcc aggccctagggaggagagtt ccagactgtt ttctctacac tgctataatt 300 ccttacactg ctgggagcagtttctttgac atactttgca actgcagagg gctttt 356 <210> SEQ ID NO 46 <211>LENGTH: 482 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:46 ttgttgaaat tttgtttgac tgctttagta caggagtata ttccccaaga caagagacct 60gagagctttt ccctggttaa gataccaagg atgatttcca aattttagac atccttcccc 120ttgttccacc aatttttttt ttcttctggg aaaatagcca ggatgattgc aaaacataag 180cttgtaaaaa ggcaaaactc catggatgta agaaagtaaa tttcttgagg gccacaccca 240tgataacgct ggaattttca tttaattcct aactcatttt ttgttgtttt tgttttttta 300aactcaaatg tgtctcttta attgaggtca cttacttggt tgggagatta atattctggt 360ggggaaactt tctttttaga gtttatattg ttttattcct tcagtcactc agtattacta 420atggggtagc ttttggaatt ttccatcccc cccactttca gattactttt ggtctttttt 480 tt482 <210> SEQ ID NO 47 <211> LENGTH: 462 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (380)<223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY: unsure <222>LOCATION: (423) <223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY:unsure <222> LOCATION: (451) <223> OTHER INFORMATION: a, c, g or t <400>SEQUENCE: 47 gggaggccct gcttcctgcg agctgtcccg gcaggacaga gactcttcccgccgcggccc 60 tgccattcca ggctgaggct gtgagcagca ccatgacaag ctccagccgcagtggctctc 120 aacagtgtgg gtctctgacc acccgacgag ctggaagtgc agaccgctgacctcccttga 180 gaacctactg ggttcttgca gtaggctcct cagcggtgtc taaacacgccactcagatga 240 ttctatgcac catcacattg gaaacttttt tcattgactg ttacttaatgagaagacttc 300 cctccgggat ggttctgaag cttccttcat aggagcaagc ctttggcgggagagcactga 360 gcagacgtgc agcatctttn ctggcttcta ccgaaacacc atggatccagacgtgggttt 420 gtngtctgca cgtggaagcc agccctgcgt ngggtgagcc tg 462 <210>SEQ ID NO 48 <211> LENGTH: 1609 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 48 atgaggctgc ggtcatcagg gctggaggag ttagaggagggcaggaagga ccttgccatt 60 ctcttatcaa ggaactgcca ggacctctgt gaagctgaggccttcagcga gctcctccca 120 ggaccgtcgg agagtcccag gaatctggct gtgctgattggtacagtctt cttcagattt 180 attctatata aagtaagcat attgtcaacc ttcctcgctcctttcaagca cctgagtcct 240 ggcatcacaa acacggagga tgacgacacc ctcagtaccagcagcgcgga ggtgaaggag 300 aaccgcaacg tgggcaacct ggccgcgcgg ccaccgccctccggggaccg ggcccggggc 360 ggcgcgcccg gcgcgaagag gaagcggccg ctggaggaggggaatggggg ccacttgtgc 420 aaactgcagc tggtctggaa gaagctgtcg tggtcggtggcgcccaagaa cgcgctggtg 480 cagctgcacg agctgaggcc gggcctgcag taccggacagtgtcgcagac gggcccggtg 540 catgccccgg tcttcgcggt agcggtggag gtgaacgggctcacgttcga gggcacaggc 600 cccaccaaga agaaggccaa gatgcgcgcg gcggagctggcactcaggtc cttcgtgcag 660 ttccccaacg cctgccaggc gcacctggcc atgggcgggggcccgggccc cggcacggac 720 ttcacctccg accaggccga tttccccgac acgctcttccaggagttcga gcccccggcg 780 ccgcgccccg gactcgcggg aggccgcccc ggggacgccgcgcttctgtc cgcggcctac 840 gggcgacggc ggctgctgtg ccgcgcgctg gacctggtgggcccgacccc cgccaccccc 900 gcggccccgg gcgagcgcaa ccccgtggtg ctgctgaaccgcctgcgcgc cgggctgcgc 960 tacgtgtgtc tggcagaacc ggccgagcgg cgcgcgcggagcttcgtgat ggccgtgagc 1020 gtggacggca ggacgttcga gggctcgggg cgcagcaagaagctggcccg gggtcaggcc 1080 gcgcaggccg cactgcagga gctgttcgac atccagatgcccggccacgc gcccggcagg 1140 gccaggagga cgccaatgcc gcagggctgg cttccacgtgcaggaccaca aaaccacgtc 1200 tggatccatg gtgtttcggt agaagccagc aaagatgctgcacgtctgct cagtgctctc 1260 ccgccaaagg cttgctccta tgaaggaagc ttcagaaccatcccggaggg aagtcttctc 1320 attaagtaac agtcaatgaa aaaagtttcc aatgtgatggtgcatagaat catctgagtg 1380 gcgtgtttag acaccgctga ggagcctact gcaagaacccagtaggttct caagggaggt 1440 cagcggtctg cacttccagc tcgtcgggtg gtcagagacccacactgttg agagccactg 1500 cggctggagc ttgtcatggt gctgctcaca gcctcagcctggaatggcag ggccgcggcg 1560 ggaagagtct ctgtcctgcc gggacagctc gcaggaagcagggcctccc 1609 <210> SEQ ID NO 49 <211> LENGTH: 272 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 49 gctcccacca cggttaaaattgagctaagt gagtatcaaa cacacttggt cactttttaa 60 agagaaagca cattacctgaaaatggcatc tttcctcttg tcaactcctg ccaaaagaaa 120 gcctcacccc ctaccccctgcacaccccag aattcatact ttcaggcagc cctctggaaa 180 ctaaggacat ctaaatctaaaggtcaggat tctgtgagag aaaaccagtc caacatgctc 240 atgaaatcct aactgtgcacagggttggat gg 272 <210> SEQ ID NO 50 <211> LENGTH: 405 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 50 ctgatcccca cctttgtcctagatgccaaa tatgcagctc tcatgggaca gccctggggt 60 ctgtgtgcaa tctgtgtccacatctgtctc ctgctagact ctgtctcact taggagtttt 120 agtactgcac agcacctggaacgagcttcc aaatccactt cttccctcca ccatctcata 180 ttaattaatc ctgccagagagggctgcaca ggccgaactg cctgagaata gcaaagaggt 240 tgtttcaggc ttgggaactacagagacacc tgtaatgggg aagggatgct cttgccaagt 300 gagccgtggg cactgctgggagccacacag gactttgcat taggtcatgc aaaccccaca 360 cagaagcagg agttggcaagggccaggcct gcagggcccc aggga 405 <210> SEQ ID NO 51 <211> LENGTH: 294<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 51gggatgcata gtgagtccaa aatcacctcc ttctcatgaa tccgggagtc cctggagctc 60acggagggct tcctggtgtc ttgaaggggt ggaatccagt ctggggcgcc ccatcttcct 120gccgcctgcg gttgctgcag ccttctgtct tcactgtgaa ccctgggata ctgcggcggt 180gctggctgga aggctggctt cccagagcag tgacccgctg tggcctgctt cctgagagct 240ctgcgtgtga cagcatttcc ttctgtctct ataaaaacac aaaaattagc cagg 294 <210>SEQ ID NO 52 <211> LENGTH: 3381 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 52 atggagaagt ttctgcagat cgcgcctcac tccctggccatcgtcctggg cccggcagag 60 gcgccggcgg gggaaaggcc aggggcagcc cggcccgcgcccccggccca gccccgccag 120 ctcgcccggc accacatcgg ctacgagatc ttcgccgacttcaaagccga gaacatgcag 180 cacttctgga acaagaaggt cacggccgcg gtggccgagaccttcttcct gggctggatc 240 gacgagcagg tcctgctgat ccagggcaag gaggaacatctggaggcgct gcgcgaaggc 300 tggacgcgcc gggccctgcg gccgccctcg ggcttccacatccgctgcct gggtgatgta 360 tcacccatca gtatgtctcc catcagtcag tctcagtttattccactcgg ggagatcctc 420 tgcttggcca tctcagcaat gaactcggca agaaagcctgtcacccaaga agcactgatg 480 gagcacctga ccacgtgctt cccaggtgtt ccaacgccaagccaagaaat tctgcggcac 540 acgctgaaca cgctggtacg ggagaggaag atctacccaactccagatgg ctacttcatc 600 gtgaccccac agacttattt cataactcct tccctcataagaactaacag taaatggtac 660 catttggacg agaggatacc tgaccggtct cagtgcacctctccgcaacc cgggaccatc 720 acgccctctg cctcaggctg tgtcagggaa aggacattgccccgaaacca ctgcgactct 780 tgccactgct gcagagaaga cgtgcacagc acgcatgcacccaccctgca aaggaagtct 840 gccaaggact gcaaagaccc ttactgtccc ccttctctgtgccaggtgcc acccactgaa 900 aagagcaaaa gtactgtaaa tttttcctat aagacagaaactctctcaaa acctaaagat 960 agtgaaaagc agtcaaaaaa attcgggcta aagttattccggttaagttt taaaaaagac 1020 aagaccaaac agctggccaa tttttctgcc cagtttcctcctgaagagtg gcccctgcga 1080 gacgaggaca cgccagctac gatccctcgg gaagtagagatggaaatcat taggcgcatt 1140 aacccagacc tgaccgtgga aaatgtcatg cggcacaccgcgctcatgaa gaaactggaa 1200 gaagaaaagg cccagaggag taaagccggg tcctctgcccatcacagcgg aaggagtaaa 1260 aagagtagga ctcatcggaa gtcccatgga aagtctcggtctcacagcaa gacacgggtg 1320 tctaaaggag acccttccga cggttcacat ctggatatcccagctgaaag agagtatgac 1380 ttttgtgatc ctcttaccag gagatccaac aaagccaaggagagatccag gtcgatggat 1440 aactccaaag gccctctggg tgcttcttct ctagggacgccggaagacct tgctgaaggc 1500 tgcagccaag acgaccagac ccccagccaa tcctacattgacgacagtac tttaaggcct 1560 gcacagaccg ttagtctcca aagggctcac atttcgtccacaagctataa agaggtgtgt 1620 attccagaga tagtcagtgg cagcaaggaa ccgtccagcgcttgcagcct tttggagcca 1680 ggaaaaccac ccgagagttt gccatcctat ggcgaactcaactcttgtcc aacaaaaaca 1740 gccacagatg actatttcca gtgcaacacc tctactatcacaagtcgagc ctgtccctcc 1800 tcaaatctca cccgaagaca cctgctgaca cattgccaggccgatgtgag aaactggaac 1860 cgtccctcgg ggacctcggc ggcacaagcc atgcctgcttcccagcgtca gcaggagtca 1920 ggagggaacc aggaagcctc ttttgactat tacaacgtctctgatgatga cgactctgag 1980 gaaggggcaa acaagaacac agaggaggag aaaaatagagaggacgtagg caccatgcag 2040 tggctcctcg agcgggagaa ggaaagagac ttgcagaggaaatttgaaaa gaacctcacc 2100 cttcttgctc caaaagaaac cgacagcagc agcaaccagagagccaccca ttcagcccgg 2160 ctcgacagca tggacagcag cagcatcaca gtggacagtggattcaactc cccacgatgc 2220 cctgcagctc tgaaggctga agcatcctac tcagaacaccaggagaaaca ggatcttaag 2280 atgccagaac aggagttggc ttgtttctta ggtcggaagatcctgaacta cgatgacatg 2340 agcaacctaa acatccccag tgagattggt gggctccctggtgtgaggct ctcgcaggct 2400 gagaggctga aagctccccg gttctccctc ttcctgggtatctatgaaca ttacagtgca 2460 acaaacttca ccatggcggc aaacagcagc aatggccagacaaagacagg ggccccactg 2520 cagcagggtg aggacccccc caagatgatt ccagtgcagttgattcgagg accaagtttg 2580 agaaaacaaa ctcagctgcc agccttgccc ctcaccgtcctcatcagcga acctgttccc 2640 ctgagaatgg cttgctcatc tatctcaggg gatacccaggcagcctctgg ctccagcgac 2700 tccctgtccc cgtcactggc cttcgaggca cagttcgtcttccctgtgaa agctgccact 2760 ctgacctccc tgctgagtat gagtgatggg gtagaatcggcgaatccttt agatctttac 2820 tgcattgttg ctgaagttat gctgattgtg actatttctgggctgtcaat ggatttttat 2880 gtggccctga gcacagtgtc tcagggaggg tcaagggtagagaatccggg tttatttatc 2940 catgcagagc agtactgtag ttacaaggtc aaaaggaaaccaccaaacaa gttaattctt 3000 catagaaaaa atagtccaaa tcctggaggg aaagtgggggtcgggggaag aagggaggca 3060 cctgacacag cgctggcaga ggctgagaca gaaggaaatgctgtcacacg cagagctctc 3120 aggaagcagg ccacagcggg tcactgctct gggaagccagccttccagcc agcaccgccg 3180 cagtatccca gggttcacag tgaagacaga aggctgcagcaaccgcaggc ggcaggaaga 3240 tggggcgccc cagactggat tccacccctt caagacaccaggaagccctc cgtgagctcc 3300 agggactccc ggattcatga gaaggaggtg attttggactcactatgcat ccccagcttc 3360 catgccagag agctagacta a 3381 <210> SEQ ID NO53 <211> LENGTH: 245 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (199) <223> OTHERINFORMATION: a, c, g or t <400> SEQUENCE: 53 gaaaacctta agatacacaggtataagatg ttattattta ctcatccatt ctgcaaatat 60 atacccctat tacctagactctgttgagga tagaggccgt gctttttccc acgtgtggag 120 tagagcaaca tagaagagtgccctgcagtg ggtgctgtaa tggagatgtg taccaggtac 180 aacaggaacc taagggggnaaaggaacccc tgagtttatc ggggggcacc agggaaggct 240 tcaca 245 <210> SEQ IDNO 54 <211> LENGTH: 388 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (282) <223> OTHERINFORMATION: a, c, g or t <400> SEQUENCE: 54 tctctatcct gacttcatgatccacccgcc ttggctccca aagtgctggg attcaggtgt 60 gagccatcat gcctgggtattttgaaaacc ttaagataca caggtataag atgttattat 120 ttactcatcc attctgcaaatatatacccc tattacctag actctgttga ggatagaggc 180 cgtgcttttt cccacgtgtggagtagagca acatagaaga gtgccctgca gtgggtgctg 240 taatggagat gtgtaccaggtacaacagga acctaagggg gnaaaggaac ccctgagttt 300 atcggggggc accagggaaggcttcacaga ggacatgatg tgagttgcca tttgaagaat 360 gagaaattgt tcttctgatgaactaaac 388 <210> SEQ ID NO 55 <211> LENGTH: 360 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 55 ccaaacagaa tctttcagaatcctgttgtt ctggcaggta ttttttaata tttccttctt 60 acaaatttcc tttttacagatgatatgcat tatagtgaca ttaacaatca atcttagaca 120 caagtgattg tttttataaataggatctcc tcaatattag tgatcctata ttaagaaaga 180 tagtacatgt gaaccaatggtaaccaaaaa gaatttgaaa agcaataatt tagtgggagc 240 tcacttggaa tataactctatgtcatcatg tatttattta agtcatattc tatgaaatat 300 cctatttgaa agcaaggacaccctttggtt gcaaccccaa gttacttatg cagtattcgt 360 <210> SEQ ID NO 56 <211>LENGTH: 1203 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 56 atcttgctct ggctaatttt cgtattttta gtagagacag ggtttcaccatgttggccag 60 tctgatctca aactcctgat ctcaagtgat ctgcctgcct tggcctcccaaagtgttggg 120 attacaggcg ccacgccagc caacacctct tttttacgaa gtactgcatgagtaacttgg 180 ggttgcaacc aaagggtgtc cttgctttca aataggatat ttcatagaatatgacttaaa 240 taaatacatg atgacataga gttatattcc aagtgagctc ccactaaattattgcttttc 300 aaattctttt tggttaccat tggttcacat gtactatctt tcttaatataggatcactaa 360 tattgaggag atcctattta taaaaacaat cacttgtgtc taagattgattgttaatgtt 420 aatataatgc atatcatctg taaaaaggaa atttgtaaga aggaaatattaaaaaatacc 480 tgccagaaca acaggattct gaaagattct gtttggaaaa aacaaacaaacgaacaaaaa 540 aacgttttat ggggctaggt tttatacctc tttcccagtt attttcttttgctttcttca 600 ccacgttgtg gcaggccagg tttcactaac tcaggcttcc ataacaacggtttcagcact 660 gaccgagtgg ttccatcaaa tattaacagc tgagagagtc agtgcccttctgcaaaggct 720 ggaatgtcac aaaagcccat caagagcttt gcctcggcct ttcctgggccttaaatcatg 780 acaggataat gaaggaattc ttaacgggac ccgtttagga gtaaataagttttattgggg 840 ggtccaaaga aactccccag gcctccacaa acaagcctta ttgggtactaaagaaactcc 900 ccaaacctcc atgatttagc aggagacaag acaaaggtat tgaccccagcacctggaccc 960 atttaggtta agtaaattta ctgaggttcc agaggaagag cttcagggctcagatcttat 1020 ttatagattt aaaaaagtga atcacttatg tctttagatg aatgtacactcacatgtaga 1080 catatagctt ataatgtaaa taagctctgg aaaactttgt agttttgagttggtcttggg 1140 atcatttcca ggctttctcc ccatacctgg ttacagaaat aaactccctcctttatcagt 1200 taa 1203 <210> SEQ ID NO 57 <211> LENGTH: 780 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:unsure <222> LOCATION: (192)..(219) <223> OTHER INFORMATION: a, c, g ort <221> NAME/KEY: unsure <222> LOCATION: (442) <223> OTHER INFORMATION:a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (452) <223> OTHERINFORMATION: a, c, g or t <400> SEQUENCE: 57 gctcgacaat tttatgactatatttaatta atatgagcac attttagagt ggagaaaaca 60 aagacttcat tcatccagcaaatatttatt cagtacccga agtgctagaa actatatgag 120 ggtagtaaat aaaatagaatattcctgtcc tactatggag ggggaatgga gagggagtgg 180 aggaatagat gnnnnnnnnnnnnnnnnnnn nnnnnnnnng ttcattgtac ataaagaagt 240 tatttttttc tgacagtaactaacaaaggt ctgggcaaga atcagagggt gaccatttta 300 agaggtggtg tttctgttgagactcaaatg ataagaagga tccagtgatg cagaaatcca 360 gggcaaggaa taggatgtttgaagcctcca tagaagaaaa gcattttata gtagatcaga 420 aagcaataac aaaaaagaaaanagaaaaaa anccatttgg caatgtctag gaacaaaaag 480 gacattaacg tgggtagaatgctgtgagct aaaaagagag tagattgaaa tgaagttaaa 540 gagaaatgga gagacagacctcatagaatt ttgccctaaa tgaaatggga agccagggaa 600 gtatgacaca gtcccataataaacctgctt ctggtgcaga atggattgga attatcaagg 660 cagttagtga ggaatccagttagaaggtga atacagtggt tcagtggtcc aggatggaaa 720 tcacagtgac ctcaactaagaaggcagcag tagaggtaga gagaagttga tagatttgtc 780 <210> SEQ ID NO 58 <211>LENGTH: 945 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:58 gctcgacaat tttatgacta tatttaatta atatgagcac attttagagt ggagaaaaca 60aagacttcat tcatccagca aatatttatt cagtacccga agtgctagaa actatatgag 120ggtagtaaat aaaatagaat attcctgtcc tactatggag ggggaatgga gagggagtgg 180aggaatagat gataaacaaa caaacaaaca agcaaactag ttcattgtac ataaagaagt 240tatttttttc tgacagtaac taacaaaggt ctgggcaaga atcagagggt gaccatttta 300agaggtggtg tttctgttga gactcaaatg ataagaagga tccagtgatg cagaaatcca 360gggcaaggaa taggatgttt gaagcctcca tagaagaaaa gcattttata gtagatcaga 420aagcaataac aaaaaagaaa aaagaaaaaa aaaacatttg gcaatgtcta ggaacaaaaa 480ggacattaac gtgggtagaa tgctgtgagc taaaaagaga gtagattgaa atgaagttaa 540agagaaatgg agagacagac ctcatagaat tttgccctaa atgaaatggg aagccaggga 600agtatgacac agtcccataa taaacctgct tctggtgcag aatggattgg aattatcaag 660gcagttagtg aggaatccag ttagaaggtg aatacagtgg ttcagtggtc caggatggaa 720atcacagtga cctcaactaa gaaggcagca gtagaggtag agagaagttg atagatttgt 780cagttaagtc ctgaatcacc ttgattgtta cctactctct ctctttggtt cttaattttg 840tcttctgtaa actgggatca attatattaa tgccaagaga tgttcagaac atgatttgag 900acaagacatg agtacctgac ataaggtagg atgcagtaat ctcag 945 <210> SEQ ID NO 59<211> LENGTH: 444 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 59 cagaatagtg aagtcctaat gcctacagga tccatttgcg acaaagaaagaattcattca 60 gataaaaagc taaccgatag gcaaagattt tcaaattatt ttcttcttaaatatttttca 120 catttgtagt gaccaaagga gacctggatt tcaatcttga ctttggacctcactagctct 180 gcagtcttga gcaaattatt taatgtcctc cgaatccgtt tcctcatctgtaagaaggga 240 tagtgtttta acttcacatg gttgagataa taatagtaaa caccttagtcttgtgtttgc 300 cacagtggta gcacatagat attcaaagtt attattccta caaaaatacggactgtactc 360 accccataat tgcattttaa aagaaagacc agttttgcaa ttttccaggattatctacat 420 gatagctttt tacacaatca gtat 444 <210> SEQ ID NO 60 <211>LENGTH: 240 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:60 cagactgtac gtcacacact cacttttgtt ttattacagg acaagtttac atagggtttg 60aataccggga ggcagggatc attagggact atcttagagt ctatttatag taccctctaa 120gttgtaatta aatttttttt tgtgatgaga tgtacacaac aatttagtat tttagccgtt 180tttaagtgta cgattcaatg acatagtcac aatgttaggc aactatcacc attgttttca 240<210> SEQ ID NO 61 <211> LENGTH: 598 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 61 gcctgggtaa gagtgagcgg agtgagacctgtctctaaaa ataaaaaacc ccaagctact 60 tatacattat atcatggtgt tatcaataagggcaattaaa aaaaaaatcc aggaacattt 120 agcttgctgt tgcggctcag gagctcttgtaaggttgcac ttaggatgtt acccagggtt 180 gaagtctgaa agctgcattg aactgaaggatttgcctccg agctcactta catggttgcc 240 gacaaggtgc tcaattgctc actatgtggacctgtccatg gagctgttca tgacatgact 300 agtttacccc aaatgagtga tccagaagaaaacgtcatgc cacaattttt ttatgaccca 360 gactgtacgt cacacactca cttttgttttattacaggac aagtttacat agggtttgaa 420 taccgggagg cagggatcat tagggactatcttagagtct atttatagta ccctctaagt 480 tgtaattaaa tttttttttg tgatgagatgtacacaacaa tttagtattt tagccgtttt 540 taagtgtacg attcaatgac atagtcacaatgttaggcaa ctatcaccat tgttttca 598 <210> SEQ ID NO 62 <211> LENGTH: 1430<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 62agggtacaac ccatagccat ccatgttcat ctttgttttg aatataattg gctagaagat 60gtacatatat ctatgtaact tcctctagca tcctccagta tggaggctgc attaagactg 120catgaaggag agggagagaa gggagaaaca gagcagctgg acaagaggac aggtataggg 180aataagggag aagccagtaa ggcaggaaag accctccgtg acaaaggggc agggaacaga 240actcaaacat ttaatggcag gtaacccagg ttagaatggt aaattgaaag gtgaatataa 300agggagaatg gtgaaatgaa ttttctgaaa ttaattgctg tgtttatagt ttttagccat 360gcatcggaat cacctcagga ctccactccc aatcaattat atatctgggg gaggaccaag 420gcgttggtat ttttcagaag ctccactggt gattctgaca gcacagctag gattaagaaa 480ctgatcaatg ggaacagcat gcctgttgca gaggagcttc cctgggaaat gtcacacaca 540gaacatcaat cttctttccc cactcctgag atccctcatt ctttggcacc aggaacagtt 600gcaattagta aaccctggtt ccctgctgtc tcacaaatcg caagagtcca acgtgtggat 660ataaactttt gttcatggga ggatctttct cccagtggaa aagcaactgg gaaaagcagg 720acacactgca cagtgactgc agtttcatcc aatgccacca cccatgcagg cataaataat 780gaacatggat gggggagtct ggagctgctg aattgtaagg ctcataaatg tttaaacttt 840ttccattaat aatatttctg ctttctgtgt atgtgtatgt agaagttctg tctttataat 900tctcaccact ttgcatcata ctttccagga ggaagaaaga acacagaaat taaaattctc 960acaaaggtta ccattaagct agaggaagac cacaccactg tgtgtccaca aagatacaga 1020gccaggccgg gttcagccat gctggtcatc tgctctatat aatacaatta tttagagatg 1080gtgggtagag aacaactaca gaaaaaaaaa aactgccaga aactagaatg tcatttttac 1140acactcattt gtagaattcc tcccagtttt tactgaaggg aagtttaaaa tgattttcat 1200ttggggaaag aactgttttg agtttaccct ataagatggc cactaaaact cacccacttt 1260catgattacc tagccatcct cagatcatct tcatgatttt cctggaaata acggaagagg 1320ccctggggat gattttattg gtagagtggg aatgtattaa aattctctac ttccttgtta 1380catggtcttt cctccaccct acaaggtgtg tgcttgtaac tcaaatttcc 1430 <210> SEQ IDNO 63 <211> LENGTH: 3120 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 63 agggtacaac ccatagccat ccatgttcat ctttgttttgaatataattg gctagaagat 60 atacatatat ctatgtaact tcctctagca tcctccagtatggaggctgc attaagactg 120 catgaaggag agggagagaa gggagaaaca gagcagctggacaagaggac aggtataggg 180 aataagggag aagccagtaa ggcaggaaag accctccgtgacaaaggggc agggaacaga 240 actcaaacat ttaatggcag gtaacccagg ttagaatggtaaattgaaag gtgaatataa 300 agggagaatg gtgaaatgaa ttttctgaaa ttaattgctgtgtttatagt ttttagccat 360 gcatcggaat cacctcagga ctccactccc aatcaattatatatctgggg gaggaccaag 420 gcgttggtat ttttcagaag ctccactggt gattctgacagcacagctag gattaagaaa 480 ctgatcaatg ggaacagcat gcctgttgca gaggagcttccctgggaaat gtcacacaca 540 gaacatcaat cttccttccc cactcctgag atccctcattctttggcacc aggaacagtt 600 gcaattagta aaccctggtt ccctgctgtc tcacaaatcgcaagagtcca acgtgtggat 660 ataaactttt gttcatggga ggatctttct cccagtggaaaagcaactgg gaaaagcagg 720 acacactgca cagtgactgc agtttcatcc aatgccaccacccatgcagg cataaataat 780 gaacatggat gggggagtct ggagctgctg aattgtaaggctcataaatg tttaaacttt 840 ttccattaat aatatttctg ctttctgtgt atgtgtatgtagaagttctg tctttataat 900 tctcaccact ttgcatcata ctttccagga ggaagaaagaacacagaaat taaaattctc 960 acaaaggtta ccattaagct agaggaagac cacaccactgtgtgtccaca aagatacaga 1020 gccaggccgg gttcagccat gctggtcatc tgctctatataatacaatta tttagagatg 1080 gtgggtagag aacaactaca gaaaaaaaaa aactgccagaaactagaatg tcatttttac 1140 acactcattt gtagaattcc tcccagtttt tactgaagggaagtttaaaa tgattttcat 1200 ttggggaaag aactgttttg agtttaccct ataagatggccactaaaact cacccacttt 1260 catgattacc tagccatcct cagatcatct tcatgattttcctggaaata acggaagagg 1320 ccctggggat gattttattg gtagagtggg aatgtattaaaattctctac ttccttgtta 1380 catggtcttt cctccaccct acaaggtgtg tgcttgtaactcaaatttcc atttgagtaa 1440 ttagcaatta ttatttaaaa ctaacctgaa aataaaaattgtattcaatt cattcatagg 1500 gagcatctac ctatttatta ttaccacata ggtgatgtgactctagaaac atcttggtat 1560 tcaaatagcc aattaaaata taaaatgtaa tgattttctaaagctactcg ttttccttct 1620 ctcatctcta tctactaatt ggataagtct attctccaaacacagcaaag atgattgaca 1680 gaattctaaa aatacacaaa tttccctatt aaagagggtgaatggatgtt agcactgtat 1740 cagacacata atattaagga gatacctgct gtttaacattaacattctgt gtagtttttg 1800 ttttgcatct ataacagaca tcaaaaagtg aataaaaacatgtcgtgaaa ttacctaaat 1860 aataaattaa cttcctggat acaaggaagt tattttagcaagttcttttt aataaaagca 1920 aagaatgggg atgtaacaat tagagaaatg tggaggaaaacataagaaag ttgggagagg 1980 gaggtcaaat ctcacttcca gttatcagta aagtgcacattcttttctgt attctgtgag 2040 gctggaggtg ctgctgatag aatgtcctca catattctgtcaatgccagg atgcaaacat 2100 cactaaataa accctcgtgt caaaatgtca catagtgattatttatgctt cttgacaaca 2160 tcaatgaatg acaaggctgc ctctacagct gtgaaaggttgcataccaat tgttagaggt 2220 aacatagatg aaggacaaag tagttatcag aattcatttaaactgtgcta ttaaattgga 2280 tatatttgca gtagccatca cagtaaatac tactgtgattctcctgaaag aatattgcct 2340 aaaaaattta acgcagaaca tttcctaatt accatgcatttctgaatctc tgctaccatg 2400 tagatattca tactgaaaga gaagtgtcat ttctttttcttctttaattt gtcattgttt 2460 cccagtgttc ttgcttctgg aattgaattt gaagtccattttttaagggt actgcaatta 2520 ttaacgaagt ctggcatgat aatgtctcag cagaatgtacacaacgtgac taagatagtt 2580 aacgccttag ttgttccaat tccatattgc ttcctgttccgaatctacta atgagtaata 2640 agagatgtag cactaatcaa tgggacaaaa gcacgcattatgaaaatact ctatcaactc 2700 catcagtaaa atttgtaagg taattataaa tttgttatcagacattttaa attgttacag 2760 tataattaaa aggtattatt ttattgggga taaccttccactctgttaat catactaaaa 2820 cacttgatgc tcaaacacat tcaaagttac cttaatcagcgtgaatttca agatcctctt 2880 aaaaatgacc tctgatacta ccctgagatt actggagtgtaaatccttgc tttgcacttt 2940 ctggctgtgt gaacttaggc aattactaaa tttgctgttcttgtattttc tcactgttaa 3000 aatgaaatag aaatagcacc tacccctgtg aggtagtttgataaaaggat taaatgagaa 3060 aatctacata aagcatagtg cctaagatct gtcacacaatattttcaaga tccatgaaac 3120 <210> SEQ ID NO 64 <211> LENGTH: 561 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 64 ctccccagatatataattga ttgggagtgg ccaaggcgtg gtatttttca gaagctccac 60 tggtgattctgacagcacag ctaggattaa gaaactgatc aatgggaaca gcatgcctgt 120 tgcagaggagcttccctggg aaatgtcaca cacagaacat caatcttcct tccccactcc 180 tgagatccctcattctttgg caccaggaac agttgcaatt agtaaaccct ggttccctgc 240 tgtctcacaaatcgcaagag tccaacgtgt ggatataaac ttttgttcat gggaggatct 300 ttctcccagtggaaaagcaa ctgggaaaag caggacacac tgcacagtga ctgcagtttc 360 atccaatgccaccacccatg caggcataaa taatgaacat ggatggggga gtctggagct 420 gctgaattgaggaagaaaga acacagaaat taaaattctc acaaaggtta ccattaagct 480 agaggaagaccacaccactg tgtgtccaca aagatacaga gccaggccgg gttcagccat 540 gctggtcatctgctctatat a 561 <210> SEQ ID NO 65 <211> LENGTH: 632 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 65 atgaattttc tgaaattaattgctgtgttt atagttttta gccatgcatc ggaatcacct 60 caggactcca ctcccaatcaattatatatc tgggggagga ccaaggcgtt ggtatttttc 120 agaagctcca ctggtgattctgacagcaca gctaggatta agaaactgat caatgggaac 180 ggcatgcctg ttgcagaggagcttccctgg gaaatgtcac acacagaaca tcaatcttcc 240 ttccccactc ctgagatccctcattctttg gcaccaggaa cagttgcaat tagtaaaccc 300 tggttccctg ctgtctcacaaatcgcaaga gtccaacgtg tggatataaa cttttgttca 360 tgggaggatc tttctcccagtggaaaagca actgggaaaa gcaggacaca ctgcacagtg 420 actgcagttt catccaatgccaccacccat gcaggcataa ataatgaaca tggatggggg 480 agtctggagc tgctgaattgaggaagaaag aacacagaaa ttaaaattct cacaaaggtt 540 accattaagc tagaggaagaccacaccact gtgtgtccac aaagatacag agccaggccg 600 ggttcagcca tgctggtcatctgctctata ta 632 <210> SEQ ID NO 66 <211> LENGTH: 398 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 66 gcagcattaa atgaatgcctgttcattcaa gcatttcatt ttgttttaaa actttaaacc 60 atttcatttc tatgacataacatcagtttt tctcatgcca agcctaaata tatatatatt 120 cttggggatc tgtgacttttctctataaga tacctctgtg tctttattcc aatgataatt 180 ttagaaatta gcatacacctatttctgcat gatcttcata actatactgt aactttctat 240 tagcataaaa tttcagtgccacctttgtag cagtttttac aacagccttg gtactgcata 300 attcatttta tgtattgacaagagatctga ttatagttat gttggcttgt ccaaatgtca 360 caattagcca ggtgtggtgtctcacacttg tgatccca 398 <210> SEQ ID NO 67 <211> LENGTH: 2487 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 67 gatgataggagttaagagag gactatagaa aactgggtct ctaagctgat gtgtcaagtc 60 acactgtcctctgcttatcc taagcttacc ttgctcaaat ttcttttttt tttctttttc 120 tttgtttttggtttttattt tttcttaaat ttcaaggata ttccttcttt tgtaaatgtc 180 acagagtatcatggctctgt cgccgaggct ggagtgcaat ggtgcagtct caggtcactg 240 caacccctgccttccaggtt caagcgattc tcctccctca gcctcccaag tagctgggat 300 tacaggcacatgccatcatg cccggctaat ttttgtattt ttggtagaga tggggtttca 360 ccatgttggccaggctggtc tggaactcct gacctcaggt gatttgccca cctcagcctc 420 ccaaagtgctgggattacag gtgtgagcca ccgtgcccgt cccaaccagg cttcttaaat 480 gaattctaagatagaaacaa caggagctgc caggactctc ttaagggctg aacctaggac 540 tgtcacagtgacatttctgc catattctgc tggtcacaag gcaagcccaa attcaaaagg 600 agagaaatagacctcttaga gtttcctaat aaaaggtaat ttcattaaaa atacaattca 660 taaattagccctatgtttac tactgtcttt tcagctcttt ttttattcca tgcattaatt 720 gattcgtcaccacttggatt gtgccaccaa tgtttctatg acatgatcta aaaaaaaaaa 780 aaaaaaaaaaaagggctcag tagttttcac ttaaaagaca aagaggccca ctgagctatt 840 acagatgttagttaggattc atttacttta atatggtaga aagaatgcta tgataccact 900 ttagtgatgaacaaaataag cttaatcaca tcctaggagc taagtattct gacattataa 960 tctcttctctcagagtccca tcacagcagt cttaggattc aagatctatt cttgggaaac 1020 attatagaaccagtgtgtca tgtacataca aatgagggaa aatataatgg ctttggtaat 1080 cctgtagttatctttcttgt catatactct ttttttcatt tttaaaaatt ggggcaaaat 1140 ttatataacataaagttaac cattctgaag tgtacaattt aatggcattt aatacattca 1200 cagtgttgtacgactttttt ttttttgagg aaaagcatat ttttaggata atgtcaaaac 1260 agattaataagatgctaata agatggccag acattcactc agaagtgttt tttgttttgt 1320 tttggtgaaatggtatgaga gatatgttgc cctacactta ggccactgca ttcccgctta 1380 agtgccaggattgtgtcagc aacaggatgg cctaaacaat ctcagtcttg tctcctgcca 1440 gccccctaaatcttccagaa ttgcaagaat aggccaggtg tggtggctca cacctgtaat 1500 cccaacactttggaaggcca aggcaggcgg atcacttgag gccaggagtt tgagaccagc 1560 ctggccaacatggcaaaccc ccatctctac taaaaataca aaaattagcc aggcaggtgt 1620 ggtggtgcatgccggtaatc tcagtttctt gggaggttcg ggtgggatga tcgcttgaac 1680 ctgggaggcggaggctgcag tgagccgaga tcacgccact gcactccagc ctgggcgaca 1740 gaatgaggctttgttctcaa aaaagtgctc aacacctgta accccaacac tttgggagcc 1800 acaggcatttggattacttg agctcaggag tttgagacca gcctgggcaa catggtgaaa 1860 tcccacctccaaccgaaaat gcaaaaacta gccgggcata gtgttatgtg cctgtggtcc 1920 cagttacttgggaagctgag atgggaggat cactagagcc caggaagtca aggttgcagt 1980 gagccatgattgcaccactg cactccaccc tgggtaacag aacgagaccc tgtctcaaaa 2040 aaaagagactaaagcagcat taaatgaatg cctgttcatt caagcatttc attttgtttt 2100 aaaactttaaaccatttcat ttctatgaca taacatcagt ttttctcatg ccaagcctaa 2160 atatatatatattcttgggg atctgtgact tttctctata agatacctct gtgtctttat 2220 tccaatgataattttagaaa ttagcataca cctatttctg catgatcttc ataactatac 2280 tgtaactttctattagcata aaatttcagt gccacctttg tagcagtttt tacaacagcc 2340 ttggtactgcataattcatt ttatgtattg acaagagatc tgattatagt tatgttggct 2400 tgtccaaatgtcacaattag ccaggtgtgg tgtctcacac ttgtgatccc aactcaggag 2460 gctgaggtgggaggattgct tgagccc 2487 <210> SEQ ID NO 68 <211> LENGTH: 1184 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 68 aaacactgcaagcagaatgg gtcaaaggag agagaggcac acaatttcag gccaggccac 60 aaagccagaaaagatctgga aaatgttact ggaggaaata caaaatcatc tggagtctaa 120 catttaaagatagggtggta gattacgaga aaactaccat acagttcatg agggtggggg 180 ggatcaaagctatatttaac aaactcggct aggagcaagc tacatattta ttctgcctgg 240 aaagtacttctggcagtgtc ttatttgctg tgtccaaaat tagtaatatc atgaataact 300 ataacttcaaaaatgaagaa tatgattctg tgttttatac tttctacatg cacttatcct 360 attatgtaactgaacggtgt ggggaaaaga gtatatctaa tatgaaagaa aaaacctctg 420 cccctgagaagtccaaattc taaaaatgac ttaatcggcc aactgtgaca aaagcaaggc 480 tttgaacatctcagtatccc tggagtctgt cttctcgtca ttcacttatt gccactgtga 540 cataccacgtgacagtctgc atggccagaa atattaatta ctgtctgggg aagatggtat 600 tatatttaatactcctttac tcctaatcct accattctcc aaaagtacaa tgaagagatg 660 ggcaggcctggggatctaat gctttcatac atcaaaaggt gacacattcc cattgccacc 720 ataagttttgctatatctgg gaagaggtca ctctttgaaa ttctatatgc aattttggga 780 aatagaatggtgttcctttt tttttaccct cctaaaaaag aatgctgaag ttattttctc 840 tcttgccaaaaagagaagaa aaaaacgaga aggagaaatc agtttgagat ttgattgcgc 900 agacgtgagaacagaggcat tgccatacca tggcagatga ctcatccagc ctttaatctt 960 ggctccaaaccctggcctct gcctgaggat tcggagaaag gcaatatgcc attctaactt 1020 gtagaccatttatttaacaa ggttttcctg acttaaacct tctctttact gcaaagctgt 1080 acattggagtatgctgattt atgaaataat aaggactttt aaagtattca agagctggag 1140 attatgtttaaaaagaaata actgttttat cagacctaaa gttt 1184 <210> SEQ ID NO 69 <211>LENGTH: 543 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:69 gtacattgaa aattgagacc tttggaggct tgtgcaagtt actgaatttg tgaagaaggt 60ctcattttcc tttcttcttt ccagagttgt tccttagtct cttcagctag tcatggctac 120tactgttctg ccctcgtgag ccggtgtttg tctgccgggc ccgccttggt cacacagtga 180aggctcattc cgtgtggtgg cttgggtgcg tctgcaccat ttggacacca agtctgtgtt 240ggtcacacag tgaaggctca ttccgtgtgg tggcttgggt gcgtctgcac catttggaca 300ccaagtctgt cttggtctca cagcgaaggc tcattccgtg tggtggcttg ggtgcgtctg 360caccatttgg acatcaagtc tgtgttggtc tcacagagaa ggctcgttct gtgcagtggc 420ttgggtgggt ctgcacacgt ggaacacacc acgtgaccag aggggcctga gtcaggccta 480cctggagtag ctcagggaag ctgtaaaggt gagaagcaaa ggcttctcca tgtgtttctg 540gga 543 <210> SEQ ID NO 70 <211> LENGTH: 560 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 70 gtacattgaa aattgagacctttggaggct tgtgcaagtt actgaatttg tgaagaaggt 60 ctcattttcc tttcttctttccagagttgt tccttagtct cttcagctag tcatggctac 120 tactgttctg ccctcgtgagccggtgtttg tctgccgggc ccgccttggt cacacagtga 180 aggctcattc cgtgtggtggcttgggtgcg tctgcaccat ttggacacca agtctgtgtt 240 ggtcacacag tgaaggctcattccgtgtgg tggcttgggt gcgtctgcac catttggaca 300 ccaagtctgt cttggtctcacagcgaaggc tcattccgtg tggtggcttg ggtgcgtctg 360 caccatttgg acatcaagtctgtgttggtc tcacagagaa ggctcgttct gtgcagtggc 420 ttgggtgggt ctgcacacgtggaacacacc acgtgaccag aggggcctga gtcaggccta 480 cctggagtag ctcagggaagctgtaaaggt gagaagcaaa ggcttctcca tgtgtttctg 540 ggacgcagaa cgcgcctaat560 <210> SEQ ID NO 71 <211> LENGTH: 546 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 71 tccaccctga tgagggaggc agatatcagaaggacactgc acatgataat gggggtgctg 60 tgagggaata atgggggccc gggccttatgcagaggtggt gtcagggaag gccttgccca 120 ggggtgacat ctgagtttag agaggtcctgagaagaggcc tcactcaggt gctctggtca 180 tcagtcccag gagcccagcc ttcgagtcatcccagcccgg gcaccagacg taagtgatga 240 ggcttccaga tgattcctgc cccagctgttcagggctccc agcagagaag agctgtaccc 300 acagagccct gcttggcttc ttgacctgcggaatccatga ccctgttaca cccctaagtt 360 ctgtgatggt tcattataac aacagatcaccggaccatgg aaattacttc tcctcttcta 420 ctctttaggc taactcgttt ttatataaacaactcatccc cacccagaca cacacacaag 480 gccctagagc agcagtttcc gaacagtgccgtatcccccg gggatctcta aaaaatactg 540 acaccc 546 <210> SEQ ID NO 72 <211>LENGTH: 676 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:72 tccaccctga tgagggaggc agatatcaga aggacactgc acatgataat gggggtgctg 60tgagggaata atgggggccc gggccttatg cagaggtggt gtcagggaag gccttgccca 120ggggtgacat ctgagtttag agaggtcctg agaagaggcc tcactcaggt gctctggtca 180tcagtcccag gagcccagcc ttcgagtcat cccagcccgg gcaccagacg taagtgatga 240ggcttccaga tgattcctgc cccagctgtt cagggctccc agcagagaag agctgtaccc 300acagagccct gcttggcttc ttgacctgcg gaatccatga ccctgttaca cccctaagtt 360ctgtgatggt tcattataac aacagatcac cggaccatgg aaattacttc tcctcttcta 420ctctttaggc taactcgttt ttatataaac aactcatccc cacccagaca cacacacaag 480gccctagagc agcagtttcc gaacagtgcc gtatcccccg gggatctcta aaaaatactg 540acacccagac aatagggggg aatcctaaaa atacctgacc aagactattc aagattgtca 600ggctcatgaa aaacaaagaa aagtctgata ctacatcctg gatggaacat aacattaaaa 660ctgagaaaat cggaaa 676 <210> SEQ ID NO 73 <211> LENGTH: 451 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 73 aaaaaagagaaaagaaaacc caggaagttg cctgataatt ctttttcatg actactgaaa 60 agatgggagcctgagagaga cctaacagga gtaaatactt taaaaacaaa ttgtatggaa 120 ctgtacccattctgaggatg actgatacag ttccagtatt caaaagaagt ataccttttc 180 cagcaaagcagatcttttgt ttacatagaa aacattttgt tatttgggaa gtttgcttaa 240 ctatttgtaagtggctcttt gcaattaatg gattaccttt ttttaatgga aagaaaataa 300 attcaaaatccataagcaaa actttagtaa tttctgactc taggatacta gcatcttcta 360 tttcttatgtgatttagact caacattttc cagtgagtta agtacttaca tcctctggct 420 gtatagctctgcccatttgc ctcgtaatac a 451 <210> SEQ ID NO 74 <211> LENGTH: 453 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 74 tgcctcatatacatttctaa ggactcctct tctcagccat ctcaagctgc ttttatcata 60 agagataggaccctgctcat tttgacaaga ggaaccatat acccagctgt gtcctccctc 120 catttgagtaaacttccaaa acttgagcca ctaaaagatt ccattgctct gaccacctaa 180 cagttctcccaacatgggtc agagagcaga acagttctct aagctggtag ggggagagaa 240 gctctatcatgttagagcag acatgaatgt gtaagggaga ataagcagag gagtgtgtgc 300 aataggcaaaccatatctct gaagtgtgat gagtataccg aggtttctag ctggaaaaat 360 gttcagggtgatgagataaa gaatatctca aagttttaat caataaatgc aaaccagcaa 420 aaatatcaataaaatatttt taatgcctac aaa 453 <210> SEQ ID NO 75 <211> LENGTH: 481 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 75 tgcctcatatacatttctaa ggactcctct tctcagccat ctcaagctgc ttttatcata 60 agagataggaccctgctcat tttgacaaga ggaaccatat acccagctgt gtcctccctc 120 catttgagtaaacttccaaa acttgagcca ctaaaagatt ccattgctct gaccacctaa 180 cagttctcccaacatgggtc agagagcaga acagttctct aagctggtag ggggagagaa 240 gctctatcatgttagagcag acatgaatgt gtaagggaga ataagcagag gagtgtgtgc 300 aataggcaaaccatatctct gaagtgtgat gagtataccg aggtttctag ctggaaaaat 360 gttcagggtgatgagataaa gaatatctca aagttttaat caataaatgc aaaccagcaa 420 aaatatcaataaaatatttt taatgcctac aaaaaaaaaa aaaaaaaaaa aattggcggc 480 c 481 <210>SEQ ID NO 76 <211> LENGTH: 492 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 76 aattgaaata aatggtctga tgctctctaa aatcttactgttacatttcc ataaagttaa 60 aaatgcaaac aaagtaagac ctccttgatt tggaacatgtcagaaagata aggaaaacag 120 ttggtatgta ttctttattt agcaaagttg taatgcacagatttgacaaa tttaaagatt 180 ttttccccta gaatataatg taactttata ataagactagagatgtacct actgcatttg 240 atacccaaaa tgtatcattt tttaatacct tcacttttatatgaaaattt actttaagta 300 atagtcatgt aataatacac agtaattatt attgtcctcattccttcttt agttttaaag 360 catttaataa ttaaacacag gaataccttt taaagtattaaaatttagta aaaatatttc 420 atctgcaaaa cagaatttgc atttaccaaa ctcaagcattataccttgca gtttgtaatt 480 gtttcactgt tt 492 <210> SEQ ID NO 77 <211>LENGTH: 291 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: unsure <222> LOCATION: (266) <223> OTHER INFORMATION: a,c, g or t <221> NAME/KEY: unsure <222> LOCATION: (268)..(269) <223>OTHER INFORMATION: a, c, g or t <221> NAME/KEY: unsure <222> LOCATION:(273)..(275) <223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY:unsure <222> LOCATION: (286)..(287) <223> OTHER INFORMATION: a, c, g ort <400> SEQUENCE: 77 tataaagtat tttcttatta cttctgctac aattttcaaaaggaattcct aggaaataca 60 cacatataaa acttaagaaa ccaaagaacc atttcaaagaatatttaatt tctttgttgg 120 tgctcaccaa attttcttat ataatttctg aaagaagtcaaatgaaaaat taagttgata 180 atgcttatag aagtatttaa aggcatttat aaattaaatacattacataa ttatcaattg 240 aataaatgtt tctaccatat gcagantnnt ttnnntttttttttgnnacg g 291 <210> SEQ ID NO 78 <211> LENGTH: 870 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 78 ggcccattca gtaaatatttcttaagactc tactttgtgt ataacaaatt gattgtcaga 60 gcttattcat aaactatcttataatggtgc aaatcaactc acaaggcctt tgtctaataa 120 aagacagctg tgctgacagctgcttgtcct cttcaacacc tgatgtgctg gaaggttcca 180 cagtaaagca cttgaaaccttgaagtcagc cctactgtct ggataagaac ccattctctc 240 ttaaactttt gaaatcaaggcttattaaat ttgattccta gaaacttaac gctagaccat 300 atttatcaag ttttaaattgataaaattta aagtttttga gaagaaaagg gggaaaatat 360 gacaaaaaca ggccttaagttccaaggaag cccagtcttc agaaacccca tttcataatc 420 acccagttgg atgaaccaaagtagctttta cctcccaagg acagagcttc tcagaaaatg 480 gggcaaaaaa tctccagacagccctactca ggttcctggt ctctgttctc atgttctgac 540 ccccagaagg cctctaaatcactcaacttg gagactcgcc agttttttct aatcagctgc 600 ctgaaagcgg ttcagagttctgtcaacaag cccttgcacg ctgggcttat taatgctggg 660 ccactcagag ccatgactcaggaacatggc ttggggtcaa cactgaaaag cagaaatcac 720 agtactgata atgggaattttgtaggagga aacaggcttt tggaacttaa tgcctttgtt 780 aggtttctgg atttgcagatctcattgtgt gggcctgctt taggaggaaa agctgggatt 840 cataacaatc taataaacctgacccaaacc 870 <210> SEQ ID NO 79 <211> LENGTH: 576 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 79 cccctcttgt atttatgatggtttttcttt ctacttgatt aagaagaaag tacaaaaagg 60 tgcttattga agcattgtcaatttgggaac aagattctca ggaatgaaat atttgtgact 120 gttctcagca tgaaaggtagagagaagatt acttgatcat tatgtgctta tttgaaagtc 180 actttcctcc catgcctcaacatctcactt tttagaatgc acactgaatg cctggaatct 240 ctttgtttta gaaaagacaaagttgcattg ttaaaaaaat aatatttctg gggaaaaggc 300 tcagtgctca acggttactctataaagaga tttaaaaaga aagtcaaggg aagataagga 360 aggtagcatg gctaacaacctcgtgggttc tactagcatt atttcatgca aaatgtcatt 420 aacctttata taggagagaataagtctggg aatcaactgg catctaaact ccatttcttg 480 ttttgtcact gcttccctggttgactatgg aaaagacatt tggtcttttg aaaccttgcc 540 cccaacaact ataacatgacttaaattcat ttatgc 576 <210> SEQ ID NO 80 <211> LENGTH: 905 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 80 ctgatcctcagttttctgtc aaagaacttc tgctcatatc caattcctat aaagtgggag 60 tactctaacctgatattacc aacattaggt tagattttat cctaatgtta ggtattagaa 120 tatatcctaacattactagt tcttcactct tactaaagtg catgttagat ttcttcaatg 180 atttttttaaattggcatgg ttgatacaaa gcatccttct atccaaatac tgtgctcagt 240 cgcctaagtgatagtttaga accatactat ctctctgtgg aaagaatgct ggggctcctg 300 gctacacactttgagtagaa gggaaaatac ccctcttgta tttatgatgg tttttctttc 360 tacttgattaagaagaaagt acaaaaaggt gcttattgaa gcattgtcaa tttgggaaca 420 agattctcaggaatgaaata tttgtgactg ttctcagcat gaaaggtaga gagaagatta 480 cttgatcattatgtgcttat ttgaaagtca ctttcctccc atgcctcaac atctcacttt 540 ttagaatgcacactgaatgc ctggaatctc tttgttttag aaaagacaaa gttgcattgt 600 taaaaaaataatatttctgg ggaaaaggct cagtgctcaa cggttactct ataaagagat 660 ttaaaaagaaagtcaaggga agataaggaa ggtagcatgg ctaacaacct cgtgggttct 720 actagcattatttcatgcaa aatgtcatta acctttatat aggagagaat aagtctggga 780 atcaactggcatctaaactc catttcttgt tttgtcactg cttccctggt tgactatgga 840 aaagacatttggtcttttga aaccttgccc ccaacaacta taacatgact taaattcatt 900 tatgc 905<210> SEQ ID NO 81 <211> LENGTH: 622 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 81 tgagatttct ttatagtcct ctgtaggtaagggttgattt tgaaattttg gctgcttgtg 60 tttaagattt gattctcatt ctcaaacatttacatgtata attttgagaa cacattttat 120 ttaacacaat atagtaaaca gaatgaatcataactatgaa cgttttccag aagcatttct 180 tagataaatt attttataaa aagaaaaacaacacaacatc taagatttag agtggaagaa 240 tatacttgta gttaacttcc ttgttgatttaaaaaatata tttggattta ttttggcagg 300 gtgggggaat catctaataa aaaatttaaagcaaacttca ttttttctaa ccagagtgaa 360 gagacaggga gagaatcaaa tatgtgtgagctccctctgt tgctctgtaa cagcatttta 420 ttcatgattt gtgatgtgat aaggaaatttttgctaatgt gtcaaaacaa attcaatttt 480 cctttaaggc aatttataac tttatttaaatggaatataa aggaagaacc ccctatatgt 540 aaaattctga ccttcaaatt tatgttaatatttttaaatt attaaaacat taaaaatgca 600 tctatttctc accactaaga gg 622 <210>SEQ ID NO 82 <211> LENGTH: 1079 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 82 gaaagatcta aatatccctg cacctcaagt agtacctatcatatgtagac accatataaa 60 tatttgtgtt gaatttaata aactcaggct gaacctactctgcatatatt gtgatgactt 120 tcagattaac ctttagacac attcacaggg ttactctcttggaagatctc tagggatcct 180 catttatttt ggtctttggg gttactgtat gttctgctgttggtgttttt tactctttga 240 gtattgtttt tcttccttta caaagatagt gtacatgtttattgttgggg gggagtcaaa 300 tcatatcaaa tgatagaaaa tgaaagccat ctccctagttctattcccca gaagagacca 360 ctgataagag tgtagtgtgt atcttttaag actattttcagcacatacat acatacgtat 420 tttgcacata tatggcataa ttaatatatt gttctaaaatttacattttc atctatctag 480 ctagctagct atatctcacg tatttccaag tgcagtattttggaagtttt aaatagtcag 540 attagggcca tgtttggtat tgtgactttg ttcctgacctgggtagagtg ctgtgcttag 600 ctgacatatt ttacaatcca gtgatcactc ctgacattatttgcagagtg ctcttaccag 660 cagaggagaa agagctactg tccccaaaga ttggagcaaatagccctgag ggaagtggaa 720 aatgtctttg gagtgttatt tcttttatct taaaatttagtgcagatctt gcattcaaag 780 acatcatggt atatctgtgt ttgtttcctt tgtttttacaaggagtttct cccaaaaaac 840 tgaacctgaa gtaatggtca ctccaggaat ttatgtcttgtttatcctat ggcttcaagt 900 accttcagtg tattgccaag tactgctcat gtacactccagctgagaaat acagtgttag 960 gtttccaaca aaagtaccta aggatttccc acagttcacttaagaaggat gcaaaagatg 1020 ttactggaat aatcatagtc gcagttagct gtagaataaaggacaggacc cggtatggt 1079 <210> SEQ ID NO 83 <211> LENGTH: 331 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 83 gtcggtattcatctcccctg ttgggaaagg tttaatctca tgggttattt cccaaaacct 60 gctttgttgctctctcattt gtaaagcatg caatatcagc tggagagagg aaggcactgg 120 aaagaatgctttggtcactt tacatcagtt ttaaagtagt ggcaaataag agaatgccta 180 ttcagggcatttattggcac ttccatgggg ggtaggtcct gaagtttcct gtggcaggtg 240 agtgaaaggcctgggaagaa ggccaaggat gaaattgatg tggagaagag gatctggctg 300 acttttccttgagaattcta agggatattt c 331 <210> SEQ ID NO 84 <211> LENGTH: 437 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:unsure <222> LOCATION: (362) <223> OTHER INFORMATION: a, c, g or t <221>NAME/KEY: unsure <222> LOCATION: (399) <223> OTHER INFORMATION: a, c, gor t <400> SEQUENCE: 84 gtattttcca gttattcact ttcactttat aaacatgacacttttgttaa gattataaaa 60 atgaattttg actgtgcatc tgcaatttta gacattttcgttatgattgg taatagaaca 120 ataaaatgtc tagccttgta aagagttaag tttacttacaagacagacat catctaattt 180 gcaacaagga ttaaagcccc actctgaatt aaaatttctgctggaaagat ttgaggcact 240 ggcaaactgt caaaggtagt attgtgcgag agtaaccagcttttgcaaga cctgtgaaag 300 taaatctctt cggccagagt ttcttaggtg tgtgaacttatgatgatcag gaatacattg 360 gnggattccc actagagaaa cgcaccatta atagttaangctttcacttt ctggtttcac 420 ttgttgccct cataaat 437 <210> SEQ ID NO 85<211> LENGTH: 526 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 85 gcagtatttc actgccacct gccaccaggg tttaaacagg ggagtagcagggagctatta 60 gggaggaggg gattgaggga tttttactaa ggcagaggca ggtgatagatttgagatttg 120 caaagtcaga acttggagcc cagttgagga atagcattac aatgttagcagcgttgttag 180 gcgagcacaa gacttcacag gtgatgctag ttcaccaaaa tgtgaacccacattaactgg 240 ttttcctttt gaatgtgatg gttccaagag gtttgcagtc agtggagatgtgtgaaaggg 300 cttggaggtg gaaatctggg taagaatgcc aagggcattc ctggtagattaaaatggtaa 360 agcaagcaac agatctggaa aagaactagg agaaactgtt agtattttctgggtgtcaac 420 gtagggaaag gcgttctaag cgttttcctt ataaataatg aaaaaatgttaaaaagccaa 480 aaaaaaagtg gggggagact gaaaattgag attatataga aaattg 526<210> SEQ ID NO 86 <211> LENGTH: 440 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION:(144)..(169) <223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY:unsure <222> LOCATION: (178)..(179) <223> OTHER INFORMATION: a, c, g ort <221> NAME/KEY: unsure <222> LOCATION: (181) <223> OTHER INFORMATION:a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (218) <223> OTHERINFORMATION: a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (403)<223> OTHER INFORMATION: a, c, g or t <400> SEQUENCE: 86 gctcgagtgttaaagtaata agtaacaaaa taattagaaa agttagaaaa ataatgacta 60 tttggcaaagatatttctca tacaatgaga aatatctttg tccaattagt ttaaaatctg 120 atgtagagaaactgtatatc tatnnnnnnn nnnnnnnnnn nnnnnnnnng atacttanna 180 nactactttagtttttagag tagttttagt ttccggcnaa aatgagcagg tacagagaac 240 atttttgtttacaatctaga caaatgtatc cattgtgtac aaattcattg aaaataatgt 300 tatattatgttatatgtgta ctcaaatact ctgggttgta attcagcaaa acactggttt 360 ttaacaagtagcttcatctt catttttgtt attttcaata aanacaaatt cttgtcatta 420 tgcaacaaggttataataaa 440 <210> SEQ ID NO 87 <211> LENGTH: 95 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 87 acaggcgtga gtgccactgtgcctgtctca ttccctcttc attattagct ggaatacttc 60 cagaaagaga catttcccttactgactgaa acaat 95 <210> SEQ ID NO 88 <211> LENGTH: 416 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 88 cgcttgtgtg gggcagggggtatatgggaa ccccattaaa ttttactgtg aatccaaaac 60 tgctctaaga aagtctggcttttttttttt ttttctgaga cagagtttca ctctgtctca 120 ggctggagtg tgatggcacgatctaggctc actgcaccct ccacttccgg gttcaagcga 180 ctctcgtgcc tcagcctctcaaagtgctag gattacaggc gtgagtgcca ctgtgcctgt 240 ctcattccct cttcattattagctggaata cttccagaaa gagacatttc ccttactgac 300 tgaaacaatt ttttgaattttatttattta tatatgagag ggagttttgc tcttgttgct 360 taggctggag tgcaatggcgtgatctcggt gcactgaagt ctccgcctcc tggatt 416 <210> SEQ ID NO 89 <211>LENGTH: 270 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:89 cttccagaca gctggccagt tatgttactg aaacagatat gtttctgaaa catacctatg 60gctatattta tacctggttt ttaaaacttt gagtcttatt ggcttcttct ggtacatttt 120ttatttgtaa tttgtcagaa tttcagtatc tgagagtata cattctgaat tttatagttt 180tagatcatgc aattacactt atttttcctt gcttttgaaa aagtaaatgt gcttcctatt 240ttcttaatga tcagttatct ttttttgttg 270 <210> SEQ ID NO 90 <211> LENGTH:148 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 90gatggatgtt gttacttagc ttgaagaagt acattaaact gcactggtct ttggcaacac 60gtcccacgtg ccatgctagg catgcaatgg attctgatct tttattgtac aagtggtgta 120aattctgatt catgacgata tgttgttg 148 <210> SEQ ID NO 91 <211> LENGTH: 853<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 91acaaatgtca tcactgatag aaaggttcag taacttactc aagaccacaa aattaggtct 60ttttgaaaat taaaaaaaat tcaaacccag gatataaact caggtttatt ttattcccaa 120gcctattaat ttcaagttac agttattatc aaactgtatg tttcttaagg caggatctga 180gttgtgtaat catttatccc catagtagct tgcccttaag aggtacttag tacatatttt 240ttgatgaatg atgttgtaca aataatggtg tcctgtataa taggttatgg tttaaaaata 300gaggaagtct ataggactct tagaaagtac ttcagatatc tgagaagact aagattgaga 360acttcttgga ggtctaattc taaattaaac ttccaagttg ggatgcaaaa ataaggagga 420tgtttggaaa catcccatga tttatattta cataagcttc ataaggagaa tgaatacaga 480ctgacctttg cttgagagcc atgtgatggc cagagaaagc ggcatgggtt aacagccaaa 540aggaaatcag cattatacta aagcagtgtt tctcaaagtg tgttccaggg aaggatatac 600ccaagaaaca atatccatca agagacttct ataaaaagag gagataattt aatacatata 660gaagcatgac atagttccac ggatccctga acaatacagg ggttgggggc attgaccctt 720gtgcagacaa aaatctgcat gtaacttttt tttgagacag agtctctctc tgcactccag 780cccgggcaca gagcaagacc ctgagacctt gtcagaaaga aataaagaga gaaagagaga 840ggagagggag agg 853 <210> SEQ ID NO 92 <211> LENGTH: 801 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure <222>LOCATION: (553)..(676) <223> OTHER INFORMATION: a, c, g or t <400>SEQUENCE: 92 cggagagaga ctctgtctca aaaataaagt aaatgaaata aatcttacgtttaaaagccc 60 tttttcatgg attttcctca gagactttgc agaaaccaag gggtcagttatttatcagtt 120 ggaacatttt ggactttatg atcacaaatg ctttttaatc tgtgtaacttcatctacaga 180 aaaccatggt ccattaagac tagagcgatc cctctatctc ttcatgccaggcttctacca 240 gggagataac ctgtacacat tactcacgag catagtgcgt aatccacatagggtaatgtc 300 tgcagatttg agtatgtgtt ccctaattct ttatctagca tgtagagtataaataacaca 360 atactggatg cttttatgga tgaacaagga ataataccta gcacctttcttctagaagtt 420 tatagtatga agagagaaga taagatgcat ctgagaaact agattaaacttgacattgtt 480 tgatcaagag ccacgtgagc aataccaaca tggttggaga cagaggagaaatccatcgtg 540 gtgaatctag agnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn 600 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn 660 nnnnnnnnnn nnnnnnccac ctctgcccca ggtacacacg gttggtataaccaaaagtat 720 ctccaggtgt tgccaaatct gatctagagc aagaaaaaac atggaacatgaaaacagtgt 780 gtttaaaata aagccagaga g 801 <210> SEQ ID NO 93 <211>LENGTH: 280 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:93 gagaacagga cccccatgct gcacagccct ggcttaacgc gggggtggcc ccagaagcgg 60gtgggcgagg ctgggcagca gggattggct gaaatcatat gcagagccca agaggcaggg 120gaaaggcggc aatttcaggg tccctttgtt cgccaggtac ctggggccca gcccgggcgg 180caggagggac tcagcccctc gcccaggcag gaagggtccc aagcagaggc ccctccctca 240ggcactcccc agcccacacc tgcagcactg ggccaagact 280 <210> SEQ ID NO 94 <211>LENGTH: 829 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: unsure <222> LOCATION: (784) <223> OTHER INFORMATION: a,c, g or t <400> SEQUENCE: 94 gaacaggacc cccatgctgc acagccctgg cttaacgcgggggtggcccc agaagcgggt 60 gggcgaggct gggcagcagg gattggctga aatcatatgcagagcccaag aggcagggga 120 aaggcggcaa tttcagggtc cctttgttcg ccaggtacctggggcccagc ccgggcggca 180 ggagggactc agcccctcgc ccaggcagga agggtcccaagcagaggccc ctccctcagg 240 cactccccag cccacacctg cagcactggg accaagactaataaaacacc cgcctcacgg 300 aagacagctt tatcttgttg atcggaagtc tgccagcccaatttatgatg gaacataaga 360 tctctaaatc tgaatttaca ctctgtagcg taacgagaggtcaataagat taaacggggg 420 ctcaggagag gaccagcgtc aggctcactg cgaggtgctgcacagaaaac ccacagccag 480 agcccctggg cccagcccag gcaagaccag aaaaggagggggcaggtggg agaccagcct 540 ggggctcccg ggaagcccac gggatggagg cgggagagccaggaggcctg gggcaaccct 600 gggacggttc ctggatcgag gagagcaggg gggtgatgagggttccctca gggctgggga 660 gccttctcct ggtctcagac ccacccccct tcagctcccaagccctgggt gcccctggct 720 ctgaggacag tgggaatctt ccctgaggca ggttcaaggacagagctctg accctgtgcc 780 aggnctgctt tgggtgccta tgaactcggt tctggctcagagcagtcct 829 <210> SEQ ID NO 95 <211> LENGTH: 170 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 95 gataaccagt aggcagcaggctactcctgg ttcgtaacat aactccacca gtgtattctg 60 gcagccgggc ttacttaagggagagcacag acattccctg ctcaaaaaca aaactgctaa 120 acgtgactcc ggtagcctccatgctctctg caagaataaa atccttgaag 170 <210> SEQ ID NO 96 <211> LENGTH:259 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 96caaatttgaa atcttaaaat ttaagaaact agtggaggaa ttggatagta catgatttca 60aaaacatgaa aactgaggac attaaatgtg caagggttag aagtttgtcg catgcaaagg 120ggaaagtgaa gatagcattt tttcacatag tttcagaagt ccagttgctg aggttaatca 180atgaaagttg tagcatcaaa ggtttaacat aaaacaactt cctaaaatca gccaggtgca 240gaggctcacg ccggtaatc 259 <210> SEQ ID NO 97 <211> LENGTH: 392 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 97 atgggtttggtcattcaact ttaagtttat gagatatatc catgttgaat tttgtagctg 60 tggtttgatgatttttacat tatatagtat tacattccat ggatagttct cagtagataa 120 tcctcctattgtttaacatt tgcgttgctt ctcattttga cctattttaa acaggcctct 180 ttaaatatacacttctctgt agtgtatgct agaaatggag tggctggaat aaaagtggct 240 gaatcatcttcaactctagt aagatgtcaa actgttttct aaaagtgttt tattttaact 300 attatatgtcaattttgaac agctctttca cttactagca atttattatc agcaacactt 360 gttattgtcagacttttaag ttttcattca ct 392 <210> SEQ ID NO 98 <211> LENGTH: 863 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:unsure <222> LOCATION: (395)..(627) <223> OTHER INFORMATION: a, c, g ort <400> SEQUENCE: 98 atgggtttgg tcattcaact ttaagtttat gagatatatccatgttgaat tttgtagctg 60 tggtttgatg atttttacat tatatagtat tacattccatggatagttct cagtagataa 120 tcctcctatt gtttaacatt tgcgttgctt ctcattttgacctattttaa acaggcctct 180 ttaaatatac acttctctgt agtgtatgct agaaatggagtggctggaat aaaagtggct 240 gaatcatctt caactctagt aagatgtcaa actgttttctaaaagtgttt tattttaact 300 attatatgtc aattttgaac agctctttca cttactagcaatttattatc agcaacactt 360 gttattgtca gacttttaag ttttcattca ctggnnnnnnnnnnnnnnnn nnnnnnnnnn 420 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn 480 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn 540 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn 600 nnnnnnnnnn nnnnnnnnnn nnnnnnngtg atcatatttatgttttgctc atttaaaaaa 660 acctggttaa atatttcaca aatcgacatt atgatatattatcttccaaa attttaataa 720 ttttgtcttt tttcacattt tagtctttag ctcagctggaattcatttct gtgtgtggtg 780 tgagataagt ctttttcatg tttttcccta tgaaataaattatttccttc tgtattgcag 840 gtagctgaga ccaatgatag ctg 863 <210> SEQ ID NO99 <211> LENGTH: 563 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 99 ggtacatctt ggctgtggat ggaaatttga catacttttt attttctttacctgttacat 60 atcaaatctt aggatgtatt acttccaagg cggttaaact tattcaagattgactgagtc 120 tcctattttc cttaaattta ctagaagtga ggctccaaga actacagaaaatagaaggaa 180 agtctccatt gagccatgaa ctgtgagcac ctggcattta agcatgaagagtagggcttc 240 tatggtaggg actggagtag gcagcattcc aggaaaggat ctcagaggtcagaaacaata 300 gattatcagt taaatacttc tggaccaaag aagaccttga aatcctggctcggtgatgta 360 tacatattta atacacaatg agaagctcct tgagggtgag tgagagtgatagggctgaag 420 aacaggagac agaagacaat tcaaatgtcc ttacacagaa gactgattatatagataatg 480 gtacattcat ataaacatga tatatttact aattaaaaca taccaacacacacaacacct 540 cgagccgcta gtctcgagtc tag 563 <210> SEQ ID NO 100 <211>LENGTH: 667 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:100 cggcaagctc taatgggaag gatacgccct gatcacacac tcctatttca aaggggtcca 60gtaccagcac ctctcaccag tggtctacac tattatacaa ctttagaaga actctggaaa 120agttttgatc tttgtgaaga ctattttaaa cctccatttg gaccatatcc tgaaaagagt 180gggaaggatt ccttggtttc catgaaatgt tcattgtttc ggttctgtcc gtggtcaaaa 240gaattgcctt tccagcctcc ggaggggagc atttcttcac acctaggatc aggagccagt 300gacagtgaga ccgaagagac ccggaaagca ctacctatac aatcattttc acatgaaaaa 360gagtctcacc aacacagaca acactcggtc ccagtcatca gtcgcccagg ttccaacgtc 420aaacccaccc tccctccaat ccctcagggc cgcaggtaga ctagcacttg atgtctgatc 480ctaacatgga aaacctgctc tgctgatgtc gaattccttg ccttacctgg ccatgggtcc 540agctgtttct cactcaaccc attacccacg gaagaatgtg tttacctgcc ttaattctat 600cagccagttt ctcttgtgat tctttggctg gtgtctttta gttttttaat taaaaaattg 660tttctta 667 <210> SEQ ID NO 101 <211> LENGTH: 3734 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 101 gagtatacat ctttttcagcactgcacaac acctattcca aaattgacca catagttgga 60 agtaaagctc tcctcagcaaatgtaaaaga acagaaatga taacaaactg tctctcagac 120 cacagtgcaa tcaaactagaactcaggatt aagaaactca ctcaaaactg ctcaactaca 180 tggaaactga acaacctgctcctgaatgac tactgtgtac ataacaaaat gaaggcagaa 240 ataaagatgt tctttgaaaccaacgagaac aaagacacaa cataccagaa tctctgggac 300 acattcaaag cagtgtgtagagggaacttt atagcactaa atgtccacaa gagaaagcag 360 gaaagatcca aaattgacaccctaatatct caattaaaag aactagaaaa gcaagagcaa 420 acacattcaa aagctagcagaaggcaagaa ataacctaaa catcagagca gaagtgaagg 480 aaatagagac acaaaaaaccttcaaaagaa ttaatgaatc caggaactgg ttttttgaaa 540 ggatcagcaa aattgatagaccgctagcaa gactaataaa gaagaaacga gagaagaatc 600 aaatagacgc aataacacacatgataaagg ggatatcacc acgatcccac agaaatacaa 660 actaccatca gagaatactataaacacttc tatgcaaata aactagaaaa tctagaagaa 720 atggataaat tcctggacacatacaccctc ccaagactaa accaggaaga agctgaatcc 780 ctgaatagac caataacagattctgaaatt gcggcaataa ttaatagcct accaaccaaa 840 aaaagtccag gaccagatggattcacacct aaattctacc agaggtataa agaggagctg 900 gtaccattcc ttctgaaattattccaatca ataacaaaag agggaatcct ccctaattca 960 ttttatgaag ccaacatcatcctgatacta aagcctggca gagacacaac aaaaaaaaga 1020 gaatttagac caatatccatgatgatcatc gatgcaaaaa tcctcagtaa aatactggca 1080 aaccaaattc agcagcacctcataaagctt atccaccacg atcaagttgg cttcatccct 1140 gggatgaaag gctggttcaacatacgcaaa tcaataaagg taatccatca tataaacaga 1200 accaaagaca aaaaccacatgattatctca atacatgcag aaaaggcctt tgacaaaatt 1260 caacagccct tcatgctaaaaactgtcaat aaactagtta ttgatgggac atatctcaaa 1320 ataataagag ctatttatgacaaacccaca gccaatatca tactgaatgg gcaaaaactg 1380 gaagcattcc ctttgagaactggaataaga cagggatgcc ctctctcacc actcctattc 1440 aacatagtgt tggaagttctggccagggca atcaggcagg agaaagaaat aaaaggtatt 1500 caattaggaa aagagaaagtcaaactgtcc ctgtttgcag atgacatgat tttatattta 1560 gaaaacccca tcgtctcagcccaaaatctc cttaagctga tgagcagctt cagcaaggtc 1620 tcaggataca aaatcaatgtgcaaaaatca caagcattcc tatacaccaa taacagacaa 1680 acagagagtc aaatgagtgaactcccattc gcaattgctt caaagagaat aaaataccta 1740 ggaatccaac ttacaagggatgtgaaggac ctcttcaagg agaactacaa accactgctc 1800 aacaaaataa aagaggacacaaacaaatgg aagaacattc catgctcatg gataggaaga 1860 atcaatattg tgaaaatggccataatgccc aaggtaattt atagattcaa tgccatcccc 1920 atcaagctac caatgactttcttcacagaa ttggaaaaaa ctactttaaa gttcatatgg 1980 aaccaaaaaa gagcccgcattgccaagaca atcctaagcc aaaagaacaa agctggaggc 2040 atcacactac ctgacttcaaactatactac aaggctacag taaccaaaac agcatggtac 2100 tggtaccaaa acagagatatagaccaatgg aacaggatag agcccttgga attaatacca 2160 cacatctaca accatctgatctttgacaaa cctgacaaaa acaagctatg gggaaaggat 2220 tccctattta ataaatggtgctgggaaaac tggctagcca tatgtagaaa gctgaaactg 2280 aatctcttcc ttacaccttatacaaaaatt aattcaagat ggattaaaga cttaaatgtt 2340 agaccgaaaa ccataaaaatcctagaaaaa aacctaggca ataccattca agacataggc 2400 gtgggcaagg acttcatgactaaaacacca aaagcaatgg caacaaaagc caaaattgac 2460 aaatgggata taattaaactaaagagcttc tgcacagcaa aagaaactac catcatagtg 2520 aacaggcaac ctacagaatgggagaaaatt tttaaaatct acccatctga caaaggacta 2580 atatccagaa tctacaaagaacttaaacaa atttacaaga aaaaaatcaa acaaccccat 2640 caaaaattgg gcaaaggatatgaacagaca cttctcaaaa gaagacattt atgcagtcaa 2700 cagacacatg aaaacatgctcatcattact ggccatcaga gaaatgcaaa tcaaaaccac 2760 aatgagatac catttcacaccagttagaat ggctagtatt aaaaagtcag gaaacaacag 2820 gggtccagta ccagcacctctcaccagtgg tctacactat tatacaactt tagaagaact 2880 ctggaaaagt tttgatctttgtgaagacta ttttaaacct ccatttggac catatcctga 2940 aaagagtggg aaggattccttggtttccat gaaatgttca ttgtttcggt tctgtccgtg 3000 gtcaaaagaa ttgcctttccagcctccgga ggggagcatt tcttcacacc taggatcagg 3060 agccagtgac agtgagaccgaagagacccg gaaagcacta cctatacaat cattttcaca 3120 tgaaaaagag tctcaccaacacagacaaca ctcggtccca gtcatcagtc gcccaggttc 3180 caacgtcaaa cccaccctccctccaatccc tcagggccgc aggtagacta gcacttgatg 3240 tctgatccta acatggaaaacctgctctgc tgatgtcgaa ttccttgcct tacctggcca 3300 tgggtccagc tgtttctcactcaacccatt acccacggaa gaatgtgttt acctgcctta 3360 attctatcag ccagtttctcttgtgattct ttggctggtg tcttttagtt ttttaattaa 3420 aaaattgttt cttacaaaaaaaacggtatt cctggaggcg attactggta cgcaatctag 3480 gcaatgctcg attcagcccactacaccctg cccctgtggg cccccagtgg ggacctttgc 3540 ggccccccct ttttgtggggggttaacgca tagccccttt taaaggatac cgttcccccc 3600 ggcctgcagg ttggcacaacaggtttaatc ctagtaaagg ggggagatcc cgctcttccc 3660 acaatcgagg ggccccgaacaaaaataacc tgtaatcaat gaggcccgac aaggccagaa 3720 cacacaagag cggc 3734<210> SEQ ID NO 102 <211> LENGTH: 353 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 102 gtggatggca gcacagattt agtgtctggttttgtcagac ccatagctgg tgaggaaaac 60 agttaaccgg aatggaaatg aaagaaagtggttcgaggtg acagcaaagc aaagaacaag 120 caaagcatag ctggtaacag aaaaaaagactgaggaatga taaacaatgg acgtaagaac 180 tccaaggcca gcaataggag gccatctagaggaagcggcc agcacacctc gatagccata 240 gggcagcgct gtttgtgcca ggaggaaaatgatgtacaga tggtgacact aggacatgaa 300 gaaagggtta ggcgatgttg agaaacctcattttaagaag acacacagtc gga 353 <210> SEQ ID NO 103 <211> LENGTH: 484<212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: unsure <222> LOCATION: (8) <223> OTHER INFORMATION: a, c, g ort <221> NAME/KEY: unsure <222> LOCATION: (11) <223> OTHER INFORMATION:a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (285) <223> OTHERINFORMATION: a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (427)<223> OTHER INFORMATION: a, c, g or t <400> SEQUENCE: 103 cggctcgnagntgttacagc tatggataat agtgttatct aagctgagtg tattgaatac 60 tttcatctagatgtcttaaa aggactttag taggatataa aaagaaaata aaaaggaatt 120 ttaaaaatcaaattaaggca aacaatctct acacattttc ccccttgtat ctatgccttt 180 acatttaggttataaagtca gcccccctcc ccagtcccac ggattagcca actatctttc 240 cgtttttgattgtntggtag tgagtactgg agaatgaatg ctgtntttaa tattatgtat 300 ttttttcataactgaaattg gcgttagaag attggattaa aaaacatgag acctaacatt 360 ttagataattgtaaatatag tgtaagcagg aagatattta ctattttcaa taataaagac 420 ataaatnacttgttttctgt attttaagaa acttttgctg gtatttgata ttttaaaaaa 480 taag 484<210> SEQ ID NO 104 <211> LENGTH: 119 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 104 gagttttact cgcatccgat gagaaaggtgtgtgtacctg ctttcatgac gatagagagc 60 agacaacttc tttctggagt ttcagcttgcttccaacagt gaaggaggaa ctgaaattt 119 <210> SEQ ID NO 105 <211> LENGTH:290 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: unsure <222> LOCATION: (19) <223> OTHER INFORMATION: a, c, gor t <400> SEQUENCE: 105 gtttaatatc tcataagcna tacacacctc gaagccatcaatgacaacct tttcttgctg 60 aatagaacag tgattgatgt catgaagaca attttatctccttttgcctt ccataatttg 120 taccagtgtt tgtcacgtgg ttgttgaata aatgaataaagaatgagaaa accagaagct 180 ctgatacata atcataatga taattatttc aatgcacaactacgggtggt gctgaactag 240 aatctatatt ttctgaaact ggctcctcta ggatctactaatgatttaaa 290 <210> SEQ ID NO 106 <211> LENGTH: 1645 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure <222>LOCATION: (60) <223> OTHER INFORMATION: a, c, g or t <400> SEQUENCE: 106tttccttgaa attaagttca ggtttgtctt tgtgtgtacc aattaatgac aagaggttan 60atagaagtat gctagatggc aaagagaaat atgttttgtg tcttcaattt tgctaaaaat 120aacccagaac atggataatt catttattaa ttgattttgg taagccaagt cctatttgga 180gaaaattaat agtttttcta aaaaagaatt ttctcaatat cacctggctt gataacattt 240ttctccttcg agttcctttt tctggagttt aacaaacttg ttctttacaa atagattata 300ttgactacca ctcactgatg ttatgatatt agtttctatt gcttactttg tatttctaat 360tttaggattc acaatttagc tggagaacta ttttttaacc tgttgcacct aaacatgatt 420gagctagaag acagttttac catatgcatg cattttctct gagttatatt ttaaaatcta 480tacatttctc ctaaatatgg aggaaatcac tggcatcaaa tgccagtctc agacggaaga 540cctaaagccc atttctggcc tggagctact tggctttgtg acctatggtg aggcataagt 600gctctgagtt tgtgttgcct cttttgtaaa atgagggttt gacttaatca gtgattttca 660tagcttaaaa tttttttgaa gaacagaact ttttttaaaa acagttagat gcaaccatat 720tatataaaac agaacagata caagtagagc taacttgcta aagaaaggat ggaggctctg 780aagctgtgac ttcattatcc cttaatactg ctatgtcctc tgtagtacct tagatttcta 840tgggacatcg tttaaaaact attgtttatg cgagagcctt gctaatttcc taaaaattgt 900ggatacattt tttctcccat gtataatttt ctcaccttct atttaaaaag aaaaaaaaag 960tcagtgtagt atttacatat tttaccctat aaggagctaa cataactttt gatttagtgt 1020tattcataaa attaggttag cagtttatta accttttgta tttgctctgg caatgtttaa 1080tatctcataa gctatacaca cctcgaagcc atcaatgaca accttttctt gctgaataga 1140acagtgattg atgtcatgaa gacaatttta tctccttttg ccttccataa tttgtaccag 1200gttatataat agtataacac tgccaaggag cggattatct catcttcatc ctgtaattcc 1260agtgtttgtc acgtggttgt tgaataaatg aataaagaat gagaaaacca gaagctctga 1320tacataatca taatgataat tatttcaatg cacaactacg ggtggtgctg aactagaatc 1380tatattttct gaaactggct cctctaggat ctactaatga tttaaatcta aaagatgaag 1440ttagtaaagc atcagaaaaa aaagtgggta ttcctacaag tcaggacatt ctacgtgact 1500acaatataat ctcacagaaa tttaacatta atacattcta agatttaatt cttagattct 1560tggtaaacaa attgctcctg tggagatgat tggcatcaca tggtgttttg agctgataca 1620cccaacactt gagctcactg caaca 1645 <210> SEQ ID NO 107 <211> LENGTH: 2241<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 107gggcctgatt tcagtttctt ccagcccttc ctattgttaa catgggggtt gtgttgaaga 60atataaagtt acaaagtcaa ggaagtagga aacattttta caagtattat gtagccatct 120tggtggagcg gtggtgaggt aggctgcaaa tgattctcct atttctttcc ctgagttcag 180aacataggaa ttagattgat agacatcaac atacccgctt tattgctgac tcatgacaac 240taatgggaag acatggctca gatgtgcagc cacagtgagc ttctgaacat ttcttctcag 300actaagctct tacacacagt tgcagttgaa agaaagaatt gcttgacatg gccacaggag 360caggcagctt cctgcagaca tgacagtcaa cgcaaactca tgtcactgtg ggcagacaca 420tgtttgcaaa gagactcaga gccaaacaag cacactcaat gtgctttgcc caaatttacc 480cattaggtaa atcttccctc ctcccaagaa gaaagtggag agagcatgag tcctcacatg 540gaaacttgaa gtcagggaaa tgaaggctca ccaattattt gtgcatgggt ccaagttttc 600cttgaaatta agttcaggtt tgtctttgtg tgtaccaatt aatgacaaga ggttagatag 660aagtatgcta gatggcaaag agaaatatgt tttgtgtctt caattttgct aaaaataacc 720cagaacatgg ataattcatt tattaattga ttttggtaag ccaagtccta tttggagaaa 780attaatagtt tttctaaaaa agaattttct caatatcacc tggcttgata acatttttct 840ccttcgagtt cctttttctg gagtttaaca aacttgttct ttacaaatag attatattga 900ctaccactca ctgatgttat gatattagtt tctattgctt actttgtatt tctaatttta 960ggattcacaa tttagctgga gaactatttt ttaacctgtt gcacctaaac atgattgagc 1020tagaagacag ttttaccata tgcatgcatt ttctctgagt tatattttaa aatctataca 1080tttctcctaa atatggagga aatcactggc atcaaatgcc agtctcagac ggaagaccta 1140aagcccattt ctggcctgga gctacttggc tttgtgacct atggtgaggc ataagtgctc 1200tgagtttgtg ttgcctcttt tgtaaaatga gggtttgact taatcagtga ttttcatagc 1260ttaaaatttt tttgaagaac agaacttttt ttaaaaacag ttagatgcaa ccatattata 1320taaaacagaa cagatacaag tagagctaac ttgctaaaga aaggatggag gctctgaagc 1380tgtgacttca ttatccctta atactgctat gtcctctgta gtaccttaga tttctatggg 1440acatcgttta aaaactattg tttatgcgag agccttgcta atttcctaaa aattgtggat 1500acattttttc tcccatgtat aattttctca ccttctattt aaaaaaaaaa aaaaagtcag 1560tgtagtattt acatatttta ccctataagg agctaacata acttttgatt tagtgttatt 1620cataaaatta ggttagcagt ttattaacct tttgtatttg ctctggcaat gtttaatatc 1680tcataagcta tacacacctc gaagccatca atgacaacct tttcttgctg aatagaacag 1740tgattgatgt catgaagaca attttatctc cttttgcctt ccataatttg taccaggtta 1800tataatagta taacactgcc aaggagcgga ttatctcatc ttcatcctgt aattccagtg 1860tttgtcacgt ggttgttgaa taaatgaata aagaatgaga aaaccagaag ctctgataca 1920taatcataat gataattatt tcaatgcaca actacgggtg gtgctgaact agaatctata 1980ttttctgaaa ctggctcctc taggatctac taatgattta aatctaaaag atgaagttag 2040taaagcatca gaaaaaaaag tgggtattcc tacaagtcag gacattctac gtgactacaa 2100tataatctca cagaaattta acattaatac attctaagat ttaattctta gattcttggt 2160aaacaaattg ctcctgtgga gatgattggc atcacatggt gttttgagct gatacaccca 2220acacttgagc tcactgcaac a 2241 <210> SEQ ID NO 108 <211> LENGTH: 437 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 108 gaataaatgaataaagaatg agaaaaccag aagctctgat acataatcat aatgataatt 60 atttcaatgcacaactacgg gtggtgctga actagaatct atattttctg aaactggctc 120 ctctaggatctactaatgat ttaaatctaa aagatgaagt tagtaaagca tcagaaaaaa 180 aaggtaaacaaattgctcct gtggagatga ttggcatcac atggtgtttt gagctgatac 240 acccaacacttgagctcact gcaacagtac cagattttca ccgctatgcc tcctttcact 300 ctgggagtcttccagaggtc ttgcactcgg gagagcatgc tcaggtttcc ccagctctac 360 aaaatcacccagaatgccaa agacttcaac acaagggtaa ataaggttga tctcagaatt 420 gtcacctcaaaaaggcc 437 <210> SEQ ID NO 109 <211> LENGTH: 2587 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 109 gaggtaggct gcaaatgattctcctatttc tttccctgag ttcagaacat aggaattaga 60 ttgatagaca tcaacatacccgctttattg ctgactcatg acaactaatg ggaagacatg 120 gctcagatgt gcagccacagtgagcttctg aacatttctt ctcagactaa gctcttacac 180 acagttgcag ttgaaagaaagaattgcttg acatggccac aggagcaggc agcttcctgc 240 agacatgaca gtcaacgcaaactcatgtca ctgtgggcag acacatgttt gcaaagagac 300 tcagagccaa acaagcacactcaatgtgct ttgcccaaat ttacccatta ggtaaatctt 360 ccctcctccc aagaagaaagtggagagagc atgagtcctc acatggaaac ttgaagtcag 420 ggaaatgaag gctcaccaattatttgtgca tgggtccaag ttttccttga aattaagttc 480 aggtttgtct ttgtgtgtaccaattaatga caagaggtta gatagaagta tgctagatgg 540 caaagagaaa tatgttttgtgtcttcaatt ttgctaaaaa taacccagaa catggataat 600 tcatttatta attgattttggtaagccaag tcctatttgg agaaaattaa tagtttttct 660 aaaaaagaat tttctcaatatcacctggct tgataacatt tttctccttc gagttccttt 720 ttctggagtt taacaaacttgttctttaca aatagattat attgactacc actcactgat 780 gttatgatat tagtttctattgcttacttt gtatttctaa ttttaggatt cacaatttag 840 ctggagaact attttttaacctgttgcacc taaacatgat tgagctagaa gacagtttta 900 ccatatgcat gcattttctctgagttatat tttaaaatct atacatttct cctaaatatg 960 gaggaaatca ctggcatcaaatgccagtct cagacggaag acctaaagcc catttctggc 1020 ctggagctac ttggctttgtgacctatggt gaggcataag tgctctgagt ttgtgttgcc 1080 tcttttgtaa aatgagggtttgacttaatc agtgattttc atagcttaaa atttttttga 1140 agaacagaac tttttttaaaaacagttaga tgcaaccata ttatataaaa cagaacagat 1200 acaagtagag ctaacttgctaaagaaagga tggaggctct gaagctgtga cttcattatc 1260 ccttaatact gctatgtcctctgtagtacc ttagatttct atgggacatc gtttaaaaac 1320 tattgtttat gcgagagccttgctaatttc ctaaaaattg tggatacatt ttttctccca 1380 tgtataattt tctcaccttctatttaaaaa aaaaaaaaaa gtcagtgtag tatttacata 1440 ttttacccta taaggagctaacataacttt tgatttagtg ttattcataa aattaggtta 1500 gcagtttatt aaccttttgtatttgctctg gcaatgttta atatctcata agctatacac 1560 acctcgaagc catcaatgacaaccttttct tgctgaatag aacagtgatt gatgtcatga 1620 agacaatttt atctccttttgccttccata atttgtacca ggttatataa tagtataaca 1680 ctgccaagga gcggattatctcatcttcat cctgtaattc cagtgtttgt cacgtggttg 1740 ttgaataaat gaataaagaatgagaaaacc agaagctctg atacataatc ataatgataa 1800 ttatttcaat gcacaactacgggtggtgct gaactagaat ctatattttc tgaaactggc 1860 tcctctagga tctactaatgatttaaatct aaaagatgaa gttagtaaag catcagaaaa 1920 aaaaggtaaa caaattgctcctgtggagat gattgggcat cacatggtgt tttgagctga 1980 tacacccaac acttgagctcactggcaaca gtaccagatt ttcaccgcta tgcctccttt 2040 cactctggga gtcttccagaggtcttgcac tcgggagagc atgctcaggt ttccccagct 2100 ctacaaaatc acccagaatgccaaagactt caacacaagg gtaaataagg ttgatctcag 2160 aattgtcacc tcaaaaaggccctgccttcc actgttcagt tctggtcatc tgcctatgag 2220 atatctgaag cttgaaagagaacacttgaa aatcactgag accgtgactc ccatcccagc 2280 acacacagca agccaagtaggttacagaga tttcttcttg ggtgatgagt tcacgccaca 2340 tggggtatgt tctccagttccagtgtgttg actccttcct gcttcccccc atcagcccct 2400 gaggtcaatg tgggcagcagcccccatggt ccaagttcta gatccgctgt ggaagacttt 2460 tttaggcaat cacacagccctgcatggagt cctgatgaga gcttgcctaa ttgttgctag 2520 gtttgtcatt ttaaatacagtgtttcttta gctagtgagt aaaattggct atataggaaa 2580 aaaaaaa 2587 <210> SEQID NO 110 <211> LENGTH: 448 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (409) <223> OTHERINFORMATION: a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (431)<223> OTHER INFORMATION: a, c, g or t <400> SEQUENCE: 110 agactacctggtatggccta aggcccccaa gtaaattaaa acattcatat caggcaggat 60 agtccaaggacttagaggtt atcttgaatc tggacacagg acaaacattt ctttggaatg 120 tgaaaagtttacacaatcaa gacttgctat gttgatcctt tactgcggta ttgttttttg 180 ttttctgacccaaaagtgat gtgcattgct tacctatcat ggccttggtt aaggtgtcag 240 tcttgcctccaaatttcctc ctaaagcatg gaattggcca taaatttgct tagtgatttt 300 ctgatttagtatcattagtt tgatgactag ttttattatg tgagtgtgat aaaaggttac 360 gcttagtatacaaagacctt cttataagtc tagttaataa aactagggnc atgtgttact 420 acaacataggngttaacttt gtctggag 448 <210> SEQ ID NO 111 <211> LENGTH: 798 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:unsure <222> LOCATION: (770) <223> OTHER INFORMATION: a, c, g or t <221>NAME/KEY: unsure <222> LOCATION: (773) <223> OTHER INFORMATION: a, c, gor t <221> NAME/KEY: unsure <222> LOCATION: (779) <223> OTHERINFORMATION: a, c, g or t <400> SEQUENCE: 111 agactacctg gtatggcctaaggcccccaa gtaaattaaa acattcatat caggcaggat 60 agtccaagga cttagaggttatcttgaatc tggacacagg acaaacattt ctttggaatg 120 tgaaaagttt acacaatcaagacttgctat gttgatcctt tactgcggta ttgttttttg 180 ttttctgacc caaaagtgatgtgcattgct tacctatcat ggccttggtt aaggtgtcag 240 tcttgcctcc aaatttcctcctaaagcatg gaattggcca taaatttgct tagtgatttt 300 ctgatttagt atcattagtttgatgactag ttttattatg tgagtgtgat aaaaggttac 360 gcttagtata caaagaccttcttataagtc tagttaataa aactagggcc atgtgttact 420 acaacatagg agttaactttgtctggaggc tttttcaagc ccaagaggtt gtcatttctt 480 tatgtgtaag atactgcccacatactatac tgaagtcagg aaccaagaac cggtcaattt 540 acctcagcca agttgcttggactgctttgc ttaataatgg gtttgccata tctactttaa 600 tgagatgtgt ggcatggtgatggttacaaa agtgatttgt taattattgc ctggtaggaa 660 ggagaacatg ttttttttgaacctatgcaa atagccacat gtctgtgaaa agtaaaggga 720 tacttttgtg taaaaaaaaaaaaaaaaaaa aaaaaaaaaa aaaaaaattn ctnggtcgnc 780 aaggcaattc gtggtcgt 798<210> SEQ ID NO 112 <211> LENGTH: 683 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (676)<223> OTHER INFORMATION: a, c, g or t <400> SEQUENCE: 112 gggaaactgggttccggagg gtcagagctg cctctgcttc aggcttcttc atgatctgga 60 tcaagcttgtttgacttcca tctacagaag tcaaccttgg cttctcaaag agcaaaatag 120 ggctgagcagactagccctg gggaaggtca ctgtgtccta aggctggggg aaccaaggga 180 agaggttggtgttatctgga tttggaagct ggaagaaggg accctacagg gctgagactc 240 aggcttctgaggagggggaa ctgcccagct aaaactggtg cctttgccag ggtacaatga 300 ggctaattctgggaacatag aaagaagctg acctggaacc agctacagcc accagtgtcc 360 aatacacagttagcaggtca tcttagagag aaagactcaa aagattgtct gtgaataccc 420 taaatccaatccaggtatac tggatgcaaa cactaggaga aaaaagaaag cttttatata 480 aggaattaaattgccctctg tctgaagaaa gaaacgatta ggaaagatgg aagtgatctg 540 aataaaatttacaaaaggag aatcagacag accaacctgg gtgatgtgtt tcaagctctg 600 ccccaggagcttgaatgtct gttccttgtg gcaggagcat tcctagagac tggtgcctca 660 ggttttatgttggtgnctct ctg 683 <210> SEQ ID NO 113 <211> LENGTH: 735 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure <222>LOCATION: (698) <223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY:unsure <222> LOCATION: (700) <223> OTHER INFORMATION: a, c, g or t <221>NAME/KEY: unsure <222> LOCATION: (704) <223> OTHER INFORMATION: a, c, gor t <221> NAME/KEY: unsure <222> LOCATION: (709) <223> OTHERINFORMATION: a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (714)<223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY: unsure <222>LOCATION: (730) <223> OTHER INFORMATION: a, c, g or t <400> SEQUENCE:113 gggaaactgg gttccggagg gtcagagctg cctctgcttc aggcttcttc atgatctgga 60tcaagcttgt ttgacttcca tctacagaag tcaaccttgg cttctcaaag agcaaaatag 120ggctgagcag actagccctg gggaaggtca ctgtgtccta aggctggggg aaccaaggga 180agaggttggt gttatctgga tttggaagct ggaagaaggg accctacagg gctgagactc 240aggcttctga ggagggggaa ctgcccagct aaaactggtg cctttgccag ggtacaatga 300ggctaattct gggaacatag aaagaagctg acctggaacc agctacagcc accagtgtcc 360aatacacagt tagcaggtca tcttagagag aaagactcaa aagattgtct gtgaataccc 420taaatccaat ccaggtatac tggatgcaaa cactaggaga aaaaagaaag cttttatata 480aggaattaaa ttgccctctg tctgaagaaa gaaacgatta ggaaagatgg aagtgatctg 540aataaaattt acaaaaggag aattcagaca gaccaacctg ggtgatgtgt ttcaagctct 600gccccaggag cttgaatgtc tgttccttgt ggcaggagca ttcctagaga ctggtgcctc 660aggttttatg ttgttgtctc tctgctccag ctcacccntn aaanctctnc cctnggatgt 720tggaactcan attct 735 <210> SEQ ID NO 114 <211> LENGTH: 601 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 114 attttgaggaatttacttgg atctttcaag atcctgctgt agctaagcaa gatcttcaag 60 aaaacccttgtcttgccttc attgtgtaaa tgcaatttgc cttgtcaaat gactaggagg 120 ccagtatagcaaggtccctt tgggaaactg ggttccggag ggtcagagct gcctctgctt 180 caggcttcttcatgatctgg atcaagcttg tttgacttcc atctacagaa gtcaaccttg 240 gcttctcaaagagcaaaata gggctgagca gactagccct ggggaaggtc actgtgtcct 300 aaggctgggggaaccaaggg aagaggttgg tgttatctgg atttggaggc tggaagaagg 360 gaccctgcagggctgagact caggcttctg aggaggggga actgcccagc tagggctgat 420 acctttgccggggtgcaatg aggctaattc tgggaatata gaaggaagct gacctggaac 480 cagctacagccaccagtgtc caatacacag ttagcaggtc atcttagaga gaaagactca 540 aaagattgtctgtgaatacc ctaaatccaa tccaggtata ctggatgcaa acactaggag 600 t 601 <210>SEQ ID NO 115 <211> LENGTH: 334 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 115 gtttgaaggg gctcaactca tgccaatata cccttgcccttgcagagtgg gacgcaagaa 60 cctgatgtta gcaaactctc cacactttaa ttcaaccctgcaaacacttt ccaagtgcct 120 tctgtttgtc aggcaatatg ccagtcacta ggaatgaagaggtagataag gatgggcctt 180 aaaatcaatc tagtgaggag ggacaacata aacaaatgaacaaacaatta tcagaaatat 240 tttttatggt gtggcaaaac agtgaaatat aggatctaatttggtttgga aagaaagaga 300 aaatttaaca gaaaaagtaa tgtttgaagc cagt 334<210> SEQ ID NO 116 <211> LENGTH: 193 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION:(38)..(90) <223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY: unsure<222> LOCATION: (94) <223> OTHER INFORMATION: a, c, g or t <221>NAME/KEY: unsure <222> LOCATION: (96)..(97) <223> OTHER INFORMATION: a,c, g or t <221> NAME/KEY: unsure <222> LOCATION: (99) <223> OTHERINFORMATION: a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (156)<223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY: unsure <222>LOCATION: (172) <223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY:unsure <222> LOCATION: (179) <223> OTHER INFORMATION: a, c, g or t <400>SEQUENCE: 116 gaatgaaaca gtgggatgca gtgaggaaga ggaaacannn nnnnnnnnnnnnnnnnnnnn 60 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn tagngnncna atgtagggggaaagttaata 120 aaaattgtat tatattaggc gttttttgtt aaatangtag atcatagctgcncttgtcnc 180 aaaaaggtaa cta 193 <210> SEQ ID NO 117 <211> LENGTH: 152<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 117ggcatgccct atgattctac gtatattaag tccaaacatc aagcagtact atctatgata 60gtaaaactgg taggctaaac attggggagt tattacattg acaacaacat gaaggggttt 120atgggacttc tggtaatgtt ttgtttcctg at 152 <210> SEQ ID NO 118 <211>LENGTH: 498 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:118 atatattttt gaatgaagtt caaggcatgt atttttatgt gttaaccaaa agatgcattc 60aaggatgtta atctcagcag gatttgtaat acaccaaaat tggaaaaaac cctcaaaatg 120tctatcaatg gtagaatgga attttataca gcaatggaaa tgcatgaact atgactatta 180gcagcaacat gaatgatttt cataaaaata gttttgagca aagaatccag atataaaaga 240gggcatgccc tatgattcta cgtatattaa gtccaaacac aagcagtact atctatgata 300gtaaaatggt aggctaaaca ttggggagtt attacattga caacaacatg aaggggttta 360tgggattctg gtaatgtttt gtttcctgat ctgggcactg ggtaccagga tgtatttact 420ttgtgaacat ttaaccatct acgatttttg gatttttctg ggtatatgtt atactttcaa 480taaaacattt tcatgaga 498 <210> SEQ ID NO 119 <211> LENGTH: 663 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 119 atgtatttgactatattatt tagttctgaa aggaattttt taatacagtg aaacattagt 60 taaaaataaatgttaactcc aaaaagaata ataaaatcat tattttttaa tcagaggctg 120 ccagtgtttttctgcttaat ctatacagca tcttagctgt aaaaatgata gaattcatgt 180 tattttactatgcattctca gtaacaataa atgtgttgct cacttctaca ttttagatag 240 ctgacataaggcctaacatg gaaagcaaaa agctatccag gaaatatttc cctgctcatg 300 catatttcctttggcataca aatcattgta aatgatggtg agttaacttc aaacatttca 360 agctacacaactaatgtaat taaaccctaa gtccaattta ctattggccc ttggtacttc 420 tgcaggctgatgttaagtgt catttttcaa gtctttcatt gcctataaac aagatggtgg 480 caattttctttattttctga acacttgcaa tttcctaaaa gtttcatact tcttcatacc 540 ttcaaatatgatttaggctt ttctagtacg taggcttccc ttttctgcct tgcttgcaga 600 ccctgttcatgcttgttaga catagcttca acgtgacatc tgggaagctt cccatgacat 660 gga 663 <210>SEQ ID NO 120 <211> LENGTH: 904 <212> TYPE: DNA <213> ORGANISM: Homosapiens <220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (684)<223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY: unsure <222>LOCATION: (888) <223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY:unsure <222> LOCATION: (893) <223> OTHER INFORMATION: a, c, g or t <221>NAME/KEY: unsure <222> LOCATION: (896) <223> OTHER INFORMATION: a, c, gor t <400> SEQUENCE: 120 aaaaaaaagg actttgagtc cattcaaagt taagtaggagctctccaggt tcttccagtg 60 acccatttac cacctctact cctcacctca catctggcttcctccagggg ccctgataca 120 gtgggtgatg ggtcctaagg gggcctccag gacccaccagccctatgagg aaagagttct 180 tcctgatcct accccttgac ttccttttct ttctcctgcaggtctcagaa cggccccgaa 240 gcctccccct gtcccctgaa ttggagagct ctccttgatgccctctgtta gggcccaccc 300 caatcccagg gcagaaggac atgagggagc aaagagcttgaggaatgcca tactccggct 360 ggtccgggac atggaaattc ggactcaggg aggacccgggctgggcaatg actgggagac 420 ttgcctgggt tcccaggact tgggggtcct gactcccagccctcatcctg cagtcccctc 480 tgttcccagc cccagccttt ctaagccatt gggaatagaatggccccttt tgttctggtg 540 tccaggggtg attgtgccaa agctcttatt tccagtgccaagcccccaga ggcttgtaag 600 agttgggatg agggatggag agggactggg tctctgggaacaggttggag gtcttatctg 660 tggactgtct gactcccagc tgangccaag atggggcatgtccccgtctc tgcttagcgt 720 ctgggtgaga aaaacaggct gtgatccaga agaagggaagatagagaagg agggaaagga 780 tgtaggcgaa ggaggtgaga gacaggatag gaggaaggaagtggaggagg aggtggtagg 840 aattggaatg aggtagaagc cgtgcagagg aagaggggagaaggacgnag gangancgat 900 gaag 904 <210> SEQ ID NO 121 <211> LENGTH:1309 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 121aaaaaaaagg actttgagtc cattcaaagt taagtaggag ctctccaggt tcttccagtg 60acccatttac cacctctact cctcacctca catctggctt cctccagggg ccctgataca 120gtgggtgatg ggtcctaagg gggcctccag gacccaccag ccctatgagg aaagagttct 180tcctgatcct accccttgac ttccttttct ttctcctgca ggtctcagaa cggccccgaa 240gcctccccct atcccctgaa ttggagagct ctccttgatg ccctctgtta gggcccaccc 300caatcccagg gcagaaggac atgagggagc aaagagcttg aggaatgcca tactccggct 360ggtccgggac atggaaattc ggactcaggg aggacccggg ctgggcaatg actgggagac 420ttgcctgggt tcccaggact tgggggtcct gactcccagc cctcatcctg cagtcccctc 480tgttcccagc cccagccttt ctaagccatt gggaatagaa tggccccttt tgttctggtg 540tccaggggtg attgtgccaa agctcttatt tccagtgcca agcccccaga ggcttgtaag 600agttgggatg agggatggag agggactggg tctctgggaa caggttggag gtcttatctg 660tggactgtct gactcccagc tgaggccaag atggggcatg tccccgtctc tgcttagcgt 720ctgggtgaga aaaacaggct gtgatccaga agaagggaag atagagaagg agggaaagga 780tgtaggcgaa ggaggtgaga gacaggatag gaggaaggaa gtggaggagg aggtggtagg 840aattggaagg aggtagaagc cgtgcagagg aagaggggag agggacgaag gaggagcgat 900gaagaagagg agggagacaa aaagagggat ggaggagaga gggagtctgg agaacaaagg 960gtcctttctc tggggagggg tgcagtgggc ggggctgaca ctgtcagcca atcctcccat 1020cggggaagag aatcctggac agggacagga tggggagggt atttataacg ggctttttgg 1080tgggagatgg gtacccagtg ggggccactg gagggtctcc gggcacactc tggcccttcc 1140cagaaagggg gtccgtcttc tcgaatcctt ccacagttgt gtattgcaaa ctacggcgca 1200ttttactatt gatcacacgt cattatcttg tcattacata ctatttctat tcaacctccc 1260ccaactgaag tgtggccgcc acaatcacca ccaaccccca cacaaccaa 1309 <210> SEQ IDNO 122 <211> LENGTH: 295 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 122 gcagtttggt aattagaaaa aaacccaaag tatgcatgcagttctgtaag ataaagtgtc 60 tgtccaggca tgcatacaac ccagcaattg catgcctgggcgcttacctt acagaaatga 120 acatttataa ttacattata atatgtacac caaattcatcacagctttat taatagaagc 180 caaactctct gtgggcttct cacagtgtac ccattgccagagtaaactgc agccttgaac 240 cattgctcag cctccttacc catgagctat gaacactgaagcaggttgca cagtg 295 <210> SEQ ID NO 123 <211> LENGTH: 714 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 123 caaaagaccctgtcaacagg attaaataca cagccaccga ctcggagaaa atattttgca 60 aaacagcgtatccggcaaag aattaatatc agaatacata atgaattctc gaaaactcga 120 agtataacaaatatcgtaca ctggaaagtg ggcaaaacca ttaataaaca tttcaccaca 180 taggatatatagatggccaa agaagcatat gaaaaagatg cgcaacatca ttagctatta 240 gggaaatgcaaattaaaacc accattagga tattagtaca gaatggttaa acatcaaaaa 300 taatagtgataacaccaaat gccaataagg aagtggagga gaaataggat cattgatata 360 ttgtttttgggaaggtaaaa tggtacagcc ctctagaaag cagtttggta attagaaaaa 420 aacccaaagtatgcatgcag ttctgtaaga taaagtgtct gtccaggcat gcatacaacc 480 cagcaattgcatgcctgggc gcttacctta cagaaatgaa catttataat tacattataa 540 tatgtacacaaaattcatca cagctttatt aatagaagcc aaactctctg tgggcttctc 600 acagtgtacccattgccaga gtaaactgca gccttgaacc attgctcagc ctccttaccc 660 atgagctatgaacactgaag caggttgcac agtgaaaaaa aaaaaaagtc gacc 714 <210> SEQ ID NO 124<211> LENGTH: 924 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 124 agagtggcct aggacagctc ctctcctgcc agagctaggc aggcgccgaagtagccgcat 60 ggccccgtca gaagacccca gggactggag agccaacctc aaaggcaccatccgtgagac 120 aggcctggag accagctccg gtgggaagct ggctggccat cagaagaccgtccccacggc 180 tcacctgact tttgttattg actgcaccca cgggaagcag ctctccctggcagcaaccgc 240 atcaccaccc caagccccca gtcccaatcg agggcttgtc accccaccaatgaagaccta 300 catcgtgttc tgtggggaaa actggcccca tctgactcgg gtgacccccatgggtggggg 360 atgccttgcc caggccaggg ccaccctgcc gctctgcaga gggtctgtggcctcagcttc 420 cttcccagtc agcccgctct gcccccagga ggttcccgag gctaaggggaaacccgtgaa 480 ggctgcgcct gtgaggtctt caacttgggg aacagtcaag gactcactgaaagccctctc 540 ctcttgtgtc tgtgggcagg ccgattagct ggaaggaccg ggctctgatgcccagaggct 600 gcaattccca gggcctggcc ctgcttcccc agctaagcag gagtcttttgtgcttgagcc 660 aaggaaacat cattagatcc gctaaggggc atctgaacaa tccgtcgagtggcagaggca 720 ggataagtca cctgcacatg aagagactca ttcattcata cagcaaatattactggtaca 780 tcttccacat gccaggccct gcaaagtgct ggggagatac catggtttacatggagctgg 840 tatttttggg gtggagggaa cccaccctga ataaataaag taacccaataaataaagaag 900 atgattttga acagcgaaaa aaaa 924 <210> SEQ ID NO 125 <211>LENGTH: 939 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:125 agagtggcct aggacagctc ctctcctgcc agagctaggc aggcgccgaa gtagccgcat 60ggccccgtca gaagacccca gggactggag agccaacctc aaaggcacca tccgtgagac 120aggcctggag accagctccg gtgggaagct ggctggccat cagaagaccg tccccacggc 180tcacctgact tttgttattg actgcaccca cgggaagcag ctctccctgg cagcaaccgc 240atcaccaccc caagccccca gtcccaatcg agggcttgtc accccaccaa tgaagaccta 300catcgtgttc tgtggggaaa actggcccca tctgactcgg gtgaccccca tgggtggggg 360atgccttgcc caggccaggg ccaccctgcc gctctgcaga gggtctgtgg cctcagcttc 420cttcccagtc agcccgctct gcccccagga ggttcccgag gctaagggga aacccgtgaa 480ggctgcgcct gtgaggtctt caacttgggg aacagtcaag gactcactga aagccctctc 540ctcttgtgtc tgtgggcagg ccgattagct ggaagggccg ggctctgatg cccagaggct 600gcaattccca gggcctggcc ctgcttcccc agctaagcag gagtcttttg tgcttgagcc 660aaggaaacat cattagatcc gctaaggggc atctgaaaca tccgtcgagt ggcagaggca 720ggataagtca cctgcacatg aagagactca ttcattcata cagcaaatat tactggtaca 780tcttccacat gccaggccct gcaaagtgct ggggagatac catggttttc ctggagctgg 840tatttttggg gtggagggaa cccaccctga ataaataaag taacccaata aataaagaag 900atgattttga acagcgaaaa aaaaattcga gctcgttgg 939 <210> SEQ ID NO 126 <211>LENGTH: 317 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:126 aaaaaggttt gaatatttaa aatcagttcc atgttagtca aagagttaca attatagttc 60aactaaacct gcagtcaatg taagtattca taccctaaga aaaagcacca caaaatgatg 120tctgtgattg ttaacggttg attggtttcc tgtgtccata gtggacaata ttatgaagca 180tagacagaaa aacatgttta ctaagaagct tttttttcct tccaggaaat tctgtaggtg 240aaacatgttg aacattgtca gttgacacat attctggtga agtctaacat taaacattaa 300actaaaaagc aagtgac 317 <210> SEQ ID NO 127 <211> LENGTH: 144 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 127 gaaaccaaatattttcctaa tgcaaaagtt gaaaccttat ctgaatggaa gttcgtagtg 60 taattccccaagttttaaat gcctgggctt cgttaatgag tttttaccaa ttgtctgcta 120 catgtgtaaaatttcacctc agca 144 <210> SEQ ID NO 128 <211> LENGTH: 161 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 128 aaccaaatattttcctaatg caaaagttga aaccttatct gaatggaagt tcgtagtgta 60 attccccaagttttaaatgc ctgggcttcg ttaatgagtt tttaccaatt gtctgctaca 120 tgtgtaaaatttcacctcag cattttgtgg ttttgttttt t 161 <210> SEQ ID NO 129 <211> LENGTH:728 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: unsure <222> LOCATION: (239) <223> OTHER INFORMATION: a, c, gor t <221> NAME/KEY: unsure <222> LOCATION: (255) <223> OTHERINFORMATION: a, c, g or t <400> SEQUENCE: 129 gcagagttag agccaatttcatttgtcctg cactgagaat gtgcattccc agcagatcat 60 ggttgagctc agcacacctgcaactcagcc agcctctttg aagggtgcac agttactaac 120 tgtcagcgtt gcacagccacctttagcagg tatgttcaga acttacattc ccaccctttc 180 taaaacagct cacagtaacaagagaacggg atttaacttt tgacatgcac actcatgana 240 accaatgatt tttgnaagccaaattgttga gaagataagg tggggattct gactactagt 300 atttttacaa atctgattgtcgttgcaggt tttgttttat tttgttgtgt taatgctgag 360 agtggagaat agattggaatatttgcctct tgtgtttctt tttgctttgt aacattgcaa 420 gtggtccaca ttttcctttatttaaaattt aaagttggtg cctggggttt ctggtgttaa 480 atagaagtga tacttctgcataaagtatta tggagatgct ctgttccatc cagggaggtg 540 caggtgaaaa gagggcagttcatcctcctc acacttaggg cagggagcag catgcagggg 600 cagatcagca gctttgcatgttgacaaatc cactctctgc tgcagatgcc taggggaagt 660 tgcagactta aattttcttttgtaaaatgg gggaacacaa acagatctta tgtcactggt 720 tactccag 728 <210> SEQID NO 130 <211> LENGTH: 680 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (375)..(563) <223>OTHER INFORMATION: a, c, g or t <221> NAME/KEY: unsure <222> LOCATION:(615) <223> OTHER INFORMATION: a, c, g or t <221> NAME/KEY: unsure <222>LOCATION: (649) <223> OTHER INFORMATION: a, c, g or t <400> SEQUENCE:130 ttccgaccta cattataaga ctgctgtaat gatctaaaac tttagctgtt ttaatatagt 60tttaaaacta atgatatctt tctctgtcag taaaatacaa actttttctt aataaaaatg 120taatggaaaa ctgttcctca tagatttttg tcactttaca aagtgacaaa atcattttgt 180tagtttatgg aaaataagct tgtaaaactt tttacctaaa agataggact gaaatttcag 240cttttttaat ttgatgatga gtttttaatt tcttttgaaa aagaatgtat gcttctaata 300atttatcaag aggaagaata ccaaaagaaa atatctgctc ttctttcttt ttacttagat 360ttttttgcat ttttnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 420nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 480nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 540nnnnnnnnnn nnnnnnnnnn nnnactgttt ttcctctgct ctcatacaac aatcaacaca 600gaaggcttct gtggnctcaa atgtggggga attttttcca cgccaagcna gcagtcaggg 660ctgcagtgtc tccaacttag 680 <210> SEQ ID NO 131 <211> LENGTH: 858 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:unsure <222> LOCATION: (375)..(563) <223> OTHER INFORMATION: a, c, g ort <400> SEQUENCE: 131 ttccgaccta cattataaga ctgctgtaat gatctaaaactttagctgtt ttaatatagt 60 tttaaaacta atgatatctt tctctgtcag taaaatacaaactttttctt aataaaaatg 120 taatggaaaa ctgttcctca tagatttttg tcactttacaaagtgacaaa atcattttgt 180 tagtttatgg aaaataagct tgtaaaactt tttacctaaaagataggact gaaatttcag 240 cttttttaat ttgatgatga gtttttaatt tcttttgaaaaagaatgtat gcttctaata 300 atttatcaag aggaagaata ccaaaagaaa atatctgctcttctttcttt ttacttagat 360 ttttttgcat ttttnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn 420 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn 480 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn 540 nnnnnnnnnn nnnnnnnnnn nnnactgttt ttcctctgctctcatacaac aatcaacaca 600 gaagacttct gtgaccaagt ggagtccttt tatgacacattccatactgt ggctgacatg 660 atgtatttct gccagatgct ggcagttgtg gaaactatcaatgcagcaat tggagtcact 720 acgtcaccgg tgctgccttc tctgatccag gtctcacatccaggtcacgc tgatgcaaaa 780 ggtttataca taaattttcg tcacctttat aaacagcgcagacggcgcta tggacaaaaa 840 aagaaaaaga tccactaa 858 <210> SEQ ID NO 132<211> LENGTH: 328 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (107)..(240) <223> OTHERINFORMATION: a, c, g or t <221> NAME/KEY: unsure <222> LOCATION:(254)..(255) <223> OTHER INFORMATION: a, c, g or t <400> SEQUENCE: 132aatttgtatg aactattttt taagatatta actcttttct cctatatttt tgttttcatt 60taaacattgt tgtgacatat tgaaatgttt atctttactt ttcattnnnn nnnnnnnnnn 120nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240tgcttttttc cttnntggtt tcttctattc cttttgagat cagtttcctt ctgttaaggg 300ctgaaatgtg gtccccctca aaaactcc 328 <210> SEQ ID NO 133 <211> LENGTH: 762<212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: unsure <222> LOCATION: (593)..(706) <223> OTHER INFORMATION:a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (748) <223> OTHERINFORMATION: a, c, g or t <221> NAME/KEY: unsure <222> LOCATION: (751)<223> OTHER INFORMATION: a, c, g or t <400> SEQUENCE: 133 cctacacactggtaatgcaa cagaatgccc aagagtgacc tcataaagca aggattccct 60 tcgtggccccttctctgctg cctctcagaa tccagacgct aaggaaaatc cctaagcaga 120 gattttctgttggatgctaa aagcaaggaa taaaagttga aaatttggaa aatgtctcaa 180 caccgtcaccagcgccactc gagagtcatt tctagttcac cagttgacac tacatcggtg 240 ggattttgcccaacattcaa gaaatttaag taaatattat ctatctccat tgcctgttaa 300 gaaatgtgctagtagaagtg tgagggcagg gtgtcagtgt tctctcagcc tcttccctca 360 gatactcgtctgcttaccaa aataagttgc atgtccttga caatctggtt tctatgattg 420 gtgaggctggcatgctatta cctttatgtg ccctgtagac ttgaatgacc agtttgacca 480 gtttgactgttagataatct gaaggctttt ctcttttttt ataatagacc ccatctcaaa 540 tcagataatgaaaattacat atcttgatat attagaaaag tatatacatt ctnnnnnnnn 600 nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 660 nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnngaga gtacacagat 720 tggcccggagtgatggtgtg cgcctgtngt nccggctgct cg 762 <210> SEQ ID NO 134 <211>LENGTH: 11677 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 134 cagccctcgg cagacggcca atggcggcgg tgctcggggc gctcggggcgacgcggcgct 60 tgttggcggc gctgcgaggc cagagcctag ggctagcggc catgtcatcaggtactcaca 120 ggttgattgc agaggagagg aaccaagcta tacttgacct taaagcagcaggatggtcgg 180 aattaagtga gagagatgcc atctacaaag aattctcctt ccacaattttaatcaggcat 240 ttggctttat gtcccgagtt gccctacaag cagagaagat gaatcatcacccagaatggt 300 tcaatgtata caacaaggtc cagataactc tcacctcaca tgactgtggtgaactgacca 360 aaaaagatgt gaagctggcc aagtttattg aaaaagcagc tgcttctgtgtgatttcttc 420 caaaatacat aagtctgaga ggctaaactt gatggctgtg ttaacatatgtcacgtgtag 480 cacagtggag aaagcaggat atggctcata atgacagtgg tgaagacctgcgaatgaagt 540 tgctagttaa cacctacatt agggtttgac ataggtctat gttatgggtcgctgcatctg 600 ctggaactca cagactttac tatagagaat caaagatccc gtatccgaagtctatggaaa 660 tgctcatggt ggtaaattcc aacagaatga aacaccaaac ttgcttaaagtaactcacgt 720 ttcaatttga aagagatatt gtcaaaattg gaggccccca ggttcctgtctgttccaaat 780 ctttgcatga tgacagtggt ttctctgatg tggtaagctt tggctttcttctgttttctt 840 tctaaaagat cactggagta gagaggagtt aaacagacat gacctttgacctcttgcatg 900 acctccacag atagcaaacc gggccgacac atggttgacg atgtccttttctacaatgaa 960 gttaatgaaa gttctgaaaa tagtgattac tttctgacat tgataggatttaggaaacct 1020 ctggataaat agcttaagca tggctgttta tgtttttgct atagacaaaaagcagcagca 1080 tgtacattgt atttggacac aagcctgcct cggttaatat attgaactattggaccacta 1140 gggttagtag ggagcggtct gtacactttc tgattcagca ttcagaaacattctaggtgg 1200 actctgtagc tttcagtttt gtaaagttat cggaaaaaca tcgggagggtttggccatca 1260 tatgtgagct ttgtgtttca atgccagtta ctcaggatta gtaaattaatgactgtccag 1320 aggacttcag ggtcaccaag ctgctgcacc tgccattggc tgactctccccggctatctg 1380 tggctgagat ggtgctgctt aggtcacgca gagcatgagc tgctgctgaaagggcacagg 1440 agatggccct tgggcttctc atcccaggat gcctgccctg cccaccaatccatgagaaga 1500 tatgtatgat ttcagtaggc cctggatcag cttgtcacct ctggtttcctgtttgctttc 1560 cactcactca gctggagttt catttccaga ctaaagtctt catcattggcttcagaaaca 1620 gcattcatct gtggctgtgc tgatgtagta caccaagaac aactgggctcttctctgtca 1680 ctttcagtgg gctaccttcc ctcacctctc caagcagcat gaaagaattctttacatttt 1740 taatctcttt tttgtttttc cctgaaagta tgctttggtg cttaaagagagaagtcacaa 1800 aagtatacta ctgagtttcc tggagatgaa atcctgttgt ccctagctatgtgaatgagc 1860 acagggatcc ctgatgccat tattttgtat attcatacgg cacacacttactgagggcct 1920 tctgtgtgcc ctaggggatt gagcacagtg acatatcagg gcaggtagaaacagatggag 1980 agctgatgcg ggctgtctta gagcagctgc cccaggaggc ccctgtggatggatgttggg 2040 caggagccct gagacgttag gggcatataa ctaaaggaca tagcaggagttataggagga 2100 gctgatccct gagggaaaca atgaagacgg agaagatggg gctaaagtttgaattgtggg 2160 gacattaatc acggtgattc ttaaaacttt gctgttgatg attttaaatggagaaaatga 2220 gtacgtaaga tgttatttcc cagttcagta tataggttgc ccacaaagtattttcctacc 2280 atgaatggtc atatatactt gttgtagaat accagggaca gcagagatggtggggtagtt 2340 acttcctttt cttacagccc aagaactttg gtgtccagga gattgaccaatttagccact 2400 gagcatttaa tacaacacag ggctacccag atcccactgt cctgatttgccctgaaagcc 2460 aaaggagtca ggagaaggtg agtggggtga atatattaat cctgagagttgaacagagca 2520 aaaatcccta ttacttttgt acttaaaaca tctctgccac atgtgctcactctttatatt 2580 ctgtttaggt ggtttatatg tgcacatccc atcctatgcc tgcagttagccaactcaggg 2640 tttatattgc ctcctttctt tttttctttt tttttttttt ttttaagagatggggtctcg 2700 ttctgtcatg cagactggag tgcagtggtg tgatcacagc tcattgtaacctccaacgcc 2760 tggactgaag tgatcctcct gccttggcct ctctggtagc tgggactacaggtgcatgcc 2820 accacaccca cctaattttt tttattttta ttttttgtag agacagtctcactatcttgc 2880 tcgggctggt cctgaactcc tgggctcaag ttatcttgct gcctcagcctcccatgggta 2940 atctttattt cctttttttt ttttttttgg agatggagtt tcgctcttgtcgcccaggct 3000 ggagtgcaat ggcacgatct tggctcactg cagtctccac ctcctgggttcaagtgattc 3060 tccatcctca gcctcctgag tagctgagat tacaggcaac tgccaccatgcgcggctaat 3120 ttatgtattt ttttttagta agagatgggg tttcaccatg ttggccagactggtcttaaa 3180 ctcctgacct caagcgacct gcctgccttg gcctcccaaa gtgctgggattacaggcatg 3240 agccgctatg cctcgtcgct gatttttatt tcttattttt tttttagagatgggggtctc 3300 actatgttgc tcaggctgat ctcaaactcc tggcctcaag tgatcctcccaccttagcct 3360 cccaagttgc tgggattata agtgtgagcc actatcccta cctcactattaccttctttg 3420 cttctcttgt tttcttttgt tctaagtcaa acccatcaca atcttttcttgtccttccag 3480 gtgttttcca gtgctgtgcc ctggatgtgc tctctttctc ttagagcccagagaacttgc 3540 ttttccccct tatatatgac ccttaacttt ttctaacaca ttattaagggcctgtgtcta 3600 tcagctgggg gcacttcttg aagggagggc ctttgtgtgg tctgtttctagtgacttcca 3660 gctttaaccc agagcctcat gattgctggg tgcccatagc ctttttgctgaatggaggca 3720 ctcagtctcc ttgggaagag agaatccatg atagacccac ttgggagctccccacttcag 3780 gggcctacac actggtaatg caacagaatg cccaagagtg acctcataaagcaaggattc 3840 ccttcgtggc cccttctctg ctgcctctca gaatccagac gctaaggaaaatccctaagc 3900 agagattttc tgttggatgc taaaagcaag gaataaaagt tgaaaatttggaaaatgtct 3960 caacaccgtc accagcgcca ctcgagagtc atttctagtt caccagttgacactacatcg 4020 gtgggatttt gcccaacatt caagaaattt aagtaaatat tatctatctccattgcctgt 4080 taagaaatgt gctagtagaa gtgtgagggc agggtgtcag tgttctctcagcctcttccc 4140 tcagatactc gtctgcttac caaaataagt tgcatgtcct tgacaatctggtttctatga 4200 ttggtgaggc tggcatgcta ttacctttat gtgccctgta gacttgaatgaccagtttga 4260 ccagtttgac tgttagataa tctgaaggct tttctctttt tttataatagaccccatctc 4320 aaatcagata atgaaaatta catatcttga tatattagaa aagtatatacattctggctg 4380 ggcgcggtgg ctcacgcctg taatcccagc actttgggag gctgagggccttagcagcgg 4440 ctgccgcagc tcaatcgcgg agcaactagc cgggcgtctg cgggagccgagcgtgggacc 4500 tgtgggccta ccacctggtg ccctcatgga gacaagaagc cctgggttgaacaacatgaa 4560 gccccagtca ctgcagctgg tactggaaga gcaggtgctg gcactacagcagcagatggc 4620 agagaatcag gcagcctcct ggcggaagct gaagaactcc caggaggcccagcagagaca 4680 agcaaccctt gtgaggaagc tgcaggccaa ggtgctgcag taccgaagctggtgccaaga 4740 gctggagaag cggctagaag ccactggagg accaatcccc cagaggtgggaaaatgtgga 4800 ggagccaaac ctggatgagc tgctggtccg attggaggag gagcaacagaggtgtgagag 4860 tctagcacag gtgaacaccc agcttcgact gcacatggaa aaagctgacgtggtgaataa 4920 agcccttagg gaagatgtgg aaaaactgac agtggactgg agccgggcccgggatgagct 4980 aatgaggaag gagagccagt ggcagatgga gcaggagtgg agtctgctgttttccttgct 5040 tgtactcaga gatctgatgg agctaaaagc tgagcatgtg aggctttcagggtctctgtt 5100 gacctgttgt ctgcgcttga ctgtgggagc acagtctcgg gaacccaacggatctggaag 5160 aatgaatggg cgggagccgg cccagctgct gctgctacta gccaagacccaggagctgga 5220 gaaggaagcc catgaaagga gccaggagtt aatacagctg aagagtcaaggggatctgga 5280 gaaggctgaa cttcaggacc gggtgaccga gctctctgct ctgttgacccagtctcagaa 5340 gcaaaatgaa gattatgaaa agatgataaa ggctctgaga gagacagtggagatcctgga 5400 gacaaatcac acagaattaa tggaacatga agcatctctt agtaggaatgcgcaagagga 5460 gaagttgtct ttacagcagg tgatcaagga tataacccag gtcatggtggaagaagggga 5520 caatatagcc caaggctctg gtcatgagaa ctctttggaa ttggactctagtatcttctc 5580 ccagtttgat taccaggatg cagacaaggc tcttactctg gtgcgttcagtgctgactcg 5640 gagacgccag gctgtgcagg acctaaggca gcagcttgca ggctgtcaagaggctgtgaa 5700 cttgttgcaa cagcagcatg atcagtggga ggaagagggc aaagccttgagacagcggct 5760 gcagaagctc actggggagc gggacactct ggcagggcag actgtggacctccagggaga 5820 ggtggactct ctcagcaagg agcgagagct gctgcagaag gccagggaagagctgcggca 5880 gcagctggag gtgctagagc aggaggcatg gcgcctgcga agggtaaatgtggagcttca 5940 gctgcagggg gactctgccc agggccagaa ggaggaacag caggaggagctgcacctggc 6000 tgtccgggag agggagcgtc ttcaggagat gctgatgggc ctggaagccaaacagtcaga 6060 atcactcagt gaactgatca ctcttcggga agccctggag tcaagtcacctggaagggga 6120 gttactgagg caagagcaaa cggaagtgac cgcagcgctg gctagggcagagcagtcaat 6180 tgcagagctg tcgagttctg aaaacaccct gaagacagaa gtagctgatcttcgggctgc 6240 agctgtcaag ctcagtgcct taaatgaggc tttggcgtta gataaagttgggctgaacca 6300 gcagcttctc cagttagagg aggagaacca gtctgtgtgc agcagaatggaggccgcaga 6360 gcaggcgaga aatgctttgc aggtcgacct ggcggaggca gagaagaggagggaagccct 6420 gtgggaaaag aacactcacc tggaggctca gctgcagaaa gctgaggaggctggggctga 6480 gctgcaggca gatctcaggg acatccaaga agagaaggaa gaaattcaaaagaaactaag 6540 tgagtcacgt caccagcagg aggcagccac gactcagctg gagcagctacatcaggaggc 6600 aaagcgacag gaagaagtgc ttgccagggc agtccaggag aaggaggccctagtacgaga 6660 gaaagcggct ctagaggtgc ggctgcaggc cgtggagcgt gaccggcaggacctcgctga 6720 acaactacag gggctcagct cagccaagga gctactggag agcagtctgtttgaagccca 6780 acaacaaaat tctgtgatag aggtcaccaa ggggcagctg gaggtccagattcaaactgt 6840 cactcaagcc aaggaagtaa tccaagggga agtgaggtgc ctgaagctggaactggacac 6900 tgaacggagt caggcagagc aggagcggga tgctgcagcc agacagctggcccaggctga 6960 gcaagaaggg aagactgcct tggagcagca gaaggcagcc catgagaaagaggtgaacca 7020 gctccgggag aaatgggaga aggagcgctc ctggcaccag caggagctggcaaaggctct 7080 ggagagctta gaaagggaaa aaatggagct ggaaatgagg ctaaaggagcagcagacaga 7140 aatggaggcc atccaggccc agagggaaga agaacggacc caggcagagagtgccctatg 7200 ccagatgcag ctggaaacag agaaggagag agtatccctc ctggagacactgctgcagac 7260 gcagaaggag ctagcagatg ccagccaaca actggaacga ctgaggcaggacatgaaagt 7320 ccagaaatta aaggagcagg agaccactgg gatactacag acccagctccaggaggctca 7380 acgggagctg aaggaggcag cccggcagca cagagatgac cttgctgccctccaagaaga 7440 gagcagctcc ctgctgcagg ataagatgga cctgcagaag caggtggaggacttgaagtc 7500 tcagctggtg gcccaggatg actcccagag gctggtggag caggaggttcaggagaagct 7560 gagagagacc caggagtata accgaattca gaaggagctg gagagagagaaagccagcct 7620 gactctctca ctgatggaaa aggaacagag actccttgtt ttacaagaagctgactctat 7680 tcgacaacaa gagctgagtg ccctgcgcca ggacatgcag gaggcccagggagaacagaa 7740 agagctcagt gctcagatgg aattactaag gcaagaggtg aaggaaaaggaggctgactt 7800 tctggcccag gaagcacagc tgctggagga gctggaggcg tctcatatcacggagcagca 7860 gctgcgagcc tccttgtggg cccaggaagc caaggcagcc caactacagctgcgactgcg 7920 cagcacagag agccagctag aagcgctggc cgcagagcag cagcccgggaaccaggccca 7980 ggcccaggcc cagctggcca gcctctactc tgccctgcag caggccctggggtctgtttg 8040 tgagagcagg cctgagctga gtggtggggg agactctgct ccttccgtctggggccttga 8100 gccagaccag aatggagcta ggagcctctt taagagaggg cccctgctgactgctctctc 8160 cgctgaggca gtagcatctg ccctccacaa gcttcatcaa gacctgtggaagactcaaca 8220 gacccgggat gttctgaggg atcaggtcca gaaactggaa gagcgtctaactgatactga 8280 ggctgagaag agccaggtcc acacagagtt gcaggatctg cagagacagctctcccagaa 8340 tcaggaagag aaatctaagt gggaaggaaa gcagaactcc ctagaatctgagctgatgga 8400 actacatgaa actatggcat ccttacagag tcgcctgcgg agagcagagctacagcgaat 8460 ggaagcccag ggtgagcgag agttacttca ggcagccaag gagaacctgacagcccaggt 8520 ggaacacctg caagcagctg tcgtagaagc cagggctcag gcaagtgctgctggcatcct 8580 ggaagaagac ctgagaacgg ctcgctcagc actgaagctg aaaaatgaggaagtagagag 8640 tgagcgtgag agagcccagg ctctgcaaga gcagggcgaa ctgaaggtggcccaagggaa 8700 ggctctgcaa gagaatttgg ccctcctgac ccagacccta gctgaaagagaagaggaggt 8760 ggagactctg cggggacaaa tccaggaact ggagaagcaa cgggaaatgcagaaggctgc 8820 tttggaattg ctgtctctgg acctgaagaa gaggaaccaa gaggtagatctgcagcaaga 8880 acagattcag gagctagaga agtgtaggtc tgttttagag catctgcccatggccgtcca 8940 ggagcgagag cagaagctga ctgtgcagag ggagcagatc agagagctcgagaaggatcg 9000 ggagactcag aggaacgtct tggagcatca gcttctagaa cttgagaagaaagaccaaat 9060 gattgagtcc cagagaggac aggttcagga cctgaaaaag cagttggttactctggaatg 9120 cctggccctg gaactggagg aaaaccatca caagatggag tgccagcaaaaactgatcaa 9180 ggagctggag ggccagaggg aaacccagag agtggctttg acccaccttacgctggacct 9240 agaagaaagg agccaggagc tgcaggcaca aagcagccag atccatgacctggagagcca 9300 cagcaccgtt ctggcaagag agctgcagga gagggaccag gaggtgaagtctcagcgaga 9360 acagatcgag gagctgcaga ggcagaaaga gcatctgact caggatctcgagaggagaga 9420 ccaggagctg atgctgcaga aggagaggat tcaggttctc gaggatcagaggacccggca 9480 gaccaagatc ctggaggagg acctggaaca gatcaagctg tccttgagagagcgaggccg 9540 ggagctgacc actcagaggc agctgatgca ggaacgggca gaggaagggaagggcccaag 9600 taaagcacag cgcgggagcc tagagcacat gaagctgatc ctgcgtgataaggagaagga 9660 ggtggaatgt cagcaggagc atatccatga actccaggag ctcaaagaccagctggagca 9720 gcagctccag ggcctgcaca ggaaggtagg tgagaccagc ctcctcctgtcccagcgaga 9780 gcaggaaata gtggtcctgc agcagcaact gcaggaagcc agggaacaaggggagctgaa 9840 ggagcagtca cttcagagtc aactggatga ggcccagaga gccctagcccagagggacca 9900 ggaactggag gctctgcagc aagaacagca gcaggcccag ggacaggaggagagggtgaa 9960 ggaaaaggca gacgccctcc agggagctct ggagcaagcc catatgacactgaaggagcg 10020 tcatggagag cttcaggacc acaaggaaca ggcacgaagg ctggaggaagagctggcagt 10080 ggagggacgg cgggtccagg ccctggagga ggtgctggga gacctaagggctgagtctcg 10140 ggaacaggag aaagctctgt tggccctcca gcagcagtgt gctgagcaggcacaggagca 10200 tgaggtggag accagggccc tgcaggacag ctggctgcag gcccaggcagtgctcaagga 10260 acgggaccag gagctggaag ctctgcgggc agaaagtcag tcctcccggcatcaggagga 10320 ggctgcccgg gcccgggctg aggctctgca ggaggccctt ggcaaggctcatgctgccct 10380 gcaggggaaa gagcagcatc tcctcgagca ggcagaattg agccgcagtctggaggccag 10440 cactgcaacc ctgcaagcct ccctggatgc ctgccaggca cacagtcggcagctggagga 10500 ggctctgagg atacaagaag gtgagatcca ggaccaggat ctccgataccaggaggatgt 10560 gcagcagctg cagcaggcac ttgcccagag ggatgaagag ctgagacatcagcaggaacg 10620 ggagcagctg ctggagaagt ctctggccca gagggtccaa gagaatatgatccaagagaa 10680 gcagaatctg gggcaagaga gagaagagga ggagataagg ggccttcatcagagtgtaag 10740 ggagctacag ctgactctag cccaaaagga acaggagatt ctggagctgagggagaccca 10800 gcaaaggaac aacctggaag ccttacccca cagccacaaa acctccccaatggaggaaca 10860 atctctaaaa cttgattctt tagagcccag gctgcagcgg gagctggagcggctacaggc 10920 agccctgaga cagacagaag ccagggagat tgagtggagg gagaaggcccaggacttggc 10980 actctcccta gcgcagacca aggccagtgt cagcagtctg caggaggtagccatgttcct 11040 acaagcctct gtcctggagc gggactcaga acagcaaagg ctgcaggatgaactggagct 11100 caccagacgg gctctggaga aggagcggct acacagccca ggtgcaaccagcacagcaga 11160 actggggtcc agaggggagc agggtgtgca gctgggagag gtctcaggagtggaggctga 11220 gcctagtcct gatggaatgg agaagcagtc atggagacaa aggcttgaacacctgcagca 11280 agcagtggcc cggctggaga ttgacaggag caggctgcag cgccacaatgtccagctgcg 11340 gagtaccttg gagcaggatg ggagaggaca gaagaactca gatgccaagtgtgtggctga 11400 actgcagaaa gaggtggtcc tgctgcaagc tcagctgact ttggagcggaagcagaagca 11460 ggactacatc acccgctcag cacagaccag ccgtgagcta gcaggcctgcaccacagcct 11520 ctcacactca cttcttgccg tggcccaggc ccctgaggcc actgtcctggaggcagagac 11580 ccgcaggctg gatgagtccc tgactcaaag tctgacatcc ccagggccagtcctgctaca 11640 ccccagcccc agcactaccc aagccgcctc caggtag 11677 <210>SEQ ID NO 135 <211> LENGTH: 862 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 135 gttcttttcc gcaatgacta gtgtcaagct catgtactcttctgattcta gactggagaa 60 gattattcaa acttgatctg tgtttcaggt ttttaaatgtcctaaaaaca gaaaattaga 120 ttcagatctc aaaaaaggaa ttttggattg actttcaaagtactaatact aattatactt 180 ttcttttggt agcgtgactc ttcttatacc taagaacatattacaaatgt caaaaccatt 240 gcattttgac attgcaaaac atgccttgaa ctcttgaactactgtgaaaa gaatcaccgt 300 tgtaaagact ttttgtaagc tagctgatac tcttaagtatgtaaaaagat tgtctttcag 360 ccgacaggcc caaaggaatg tatataagga aggaatatgaaaaaataaat taggttttaa 420 aataggaatt gggcaataaa ctgtatcaaa aatatgtagatggattttag tagttgtaat 480 ttaaatgtgg aaggtgaaga gaatttcaaa ctccaaagagaaatgaatga tattcagatg 540 tttcattaat ttctagtctg tgaaaatatg cattttatagtaatatgtat agacttattt 600 tatttagaaa taatagtgtt ttagaattta ttaaaaactcagtgatagcc tttataccaa 660 aatgtttaac tttaccaaca gcaagtcata aaagtatttattttaaagct ttttaatatt 720 atcgtgtaac tttcatctgt cttcagatgt aaataattatctgcctaaat gttatatttt 780 tatgtatgca ttttctgaaa atgtattgtt ttgtaaagtgggaaagataa taaatcaagc 840 acttcttgca cttgtttctg tg 862 <210> SEQ ID NO136 <211> LENGTH: 1026 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 136 gccctttttt tctctttata caaaatgaga gtatctgagccaaaatatta aattctagtt 60 cttttccgca atgactagtg tcaagctcat gtactcttctgattctagac tggagaagat 120 tattcaaact tgatctgtgt ttcaggtttt taaatgtcctaaaaacagaa aattagattc 180 agatctcaaa aaaggaattt tggattgact ttcaaagtactaatactaat tatacttttc 240 ttttggtagc gtgactcttc ttatacctaa gaacatattacaaatgtcaa aaccattgca 300 ttttgacatt gcaaaacatg ccttgaactc ttgaactactgtgaaaagaa tcaccgttgt 360 aaagactttt tgtaagctag ctgatactct taagtatgtaaaaagattgt ctttcagccg 420 acaggcccaa aggaatgtat ataaggaagg aatatgaaaaaataaattag gttttaaaat 480 aggaattggg caataaactg tatcaaaaat atgtagatggattttagtag ttgtaattta 540 aatgtggaag gtgaagagaa tttcaaactc caaagagaaatgaatgatat tcagatgttt 600 cattaatttc tagtctgtga aaatatgcat tttatagtaatatgtataga cttattttat 660 ttagaaataa tagtgtttta gaatttatta aaaactcagtgatagccttt ataccaaaat 720 gtttaacttt accaacagca agtcataaaa gtatttattttaaagctttt taatattatc 780 gtgtaacttt catctgtctt cagatgtaaa taattatctgcctaaatgtt atatttttat 840 gtatgcattt tctgaaaatg tattgttttg taaagtgggaaagataataa atcaagcact 900 tcttgcactt gtttctgtga agcatataga actctattttaaataaggac gatgtgtcgt 960 acaacaacaa atctacttgc ccgtcggttt cccggctgcgatctggccct tgccgtacca 1020 catttc 1026 <210> SEQ ID NO 137 <211> LENGTH:611 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 137gaattttaat agggccacta agaatctgag tgctttagga gattaccctt atacccactg 60ccatcacatc cagtcaggcc tgttgtgctc tatataaatc ttcccagctg aggggcaggt 120gcgggctaaa atccaactgg caattggctc ccagacataa ttttatattt tacagagaag 180catcttattg gcttatatgt gtttaaagaa tggtctggct tatacatctt cagaaaatga 240gaattaaaaa gtcaaaataa ttcttgacat ctacagattg aacaaagaac ttagaagaaa 300taatacttta tcttttcatc ctggcattcc tgagagaaga gaaattgatt gtttatcatg 360ttggtttaat ttttcaaccc agacaatctg cagcaaggca catggacccc aattttgata 420tcgtccatac agttttcatt ctatgcatgg agctaattac tgactttgcc tgtaaagaga 480ggattgtgtg cctaaatttt gtctaacaaa tgcaagcgta gaatgacatt tactaatatt 540tctatttctt ccataggcta aataatagta actaagtatt tttaaggaca cagccctttt 600tttctcttta t 611 <210> SEQ ID NO 138 <211> LENGTH: 787 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 138 gagaaacaaa agctaattagccttgggtgt ctgcttagcc ccgtggttgg agcaggagct 60 cttgacttcc ggtcttggaactctcgccaa tagtgtgtgg aattgcactc tcacacctgc 120 tgggagacct tgcttttcaagggggtgtac agctcctcac cccagaaaaa tttgagatct 180 actggtttgg agggaaatgggagataagaa tcttatgact taaggaaaac ttattttttg 240 ttatcaaatt atagtgttccccattgaaaa tttcctttaa aactgtaaag atctttagga 300 agtagagagt ggaagccgatgctgttctgt tcccgagggt cacctgcagc tgttcccgag 360 ggtcacctcc agctgttcccgagagtcacc tccagctgtt cccgagggtc acctccagct 420 ggttgggtgc atgttcttacacaagattgt aagagacgat ctatgctgtt ctgcctgtat 480 tacttgggag gcctttggctcaaatgatgg aaaactctgc cccaaactca ctaatgaata 540 aagaaatgga tcatctcatggatgagggcg tccagaggac aagggtggcc ctggggcagt 600 ggttggttgc tgctgtcatccaggacttgg gttccgtcct ctgccctctg ccaccctctg 660 tgctggcgtc aaggtggcaaggtgtgagtt tccccgagtc acatcagctg cgacaaaatc 720 cagaggcagg aaagacctagagttcttcta aacaatgagg aaattaacat gtaaccagct 780 agaagtg 787 <210> SEQ IDNO 139 <211> LENGTH: 927 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (568)..(738) <223>OTHER INFORMATION: a, c, g or t <400> SEQUENCE: 139 tcaaagtgtatagtaaatat ctaaacaaat gaaagggaca agatatagaa ggaatcttag 60 gatcagctgagagataattg aatactttcc taaaagaaca caatactgga agggatgggg 120 ctttgtgggacaattgctat tttgaattct taggtgtcca actttacaac caaggtttac 180 aaatattttaaatggtgatt tagtcagcag aagggaagac tcaaatagaa cataattagc 240 ttaagcttacctctagttgt agagtataca ggttttgacc tcaaaatttg aaaaatcgca 300 atttttatctaagtgcaatc aagttttcct tatttgggga tggccataat tgtctctcat 360 ggcaactcaactgtatgcaa cattgaacca tcctttgttg caaaggaacc tgcctgggcc 420 tgcttcctgttagataaatg gtcccaactg gcataactta caagtttggt acataaacta 480 tctgattacaatactaacgc tatacgtgag caggtagtaa aatatgggca tttacatcat 540 ttattcaccaagaactgtaa ttaggcannn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 600 nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 660 nnnnnnnnnnnnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 720 nnnnnnnnnnnnnnnnnngc tcttggctag tttctgtttc ccactggtgt tagggtttta 780 acttctgttgattcacagga tgcaggtggg ggagagatgg caagggggaa aatatgaatt 840 tgattactcaagtggaacaa ggtgaaacca ttagtttaaa ataattttag gctgaaaaat 900 taaaatcattaaattatttc cttcatg 927 <210> SEQ ID NO 140 <211> LENGTH: 4651 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 140 gaaacttgaaaaaaaaagac agttaatcct cattatttgt ggattctgta tttgtgaatt 60 tgcctgctcgttaaaattta gaacacccca gtggatcctc ggtgcttctg tggtcatggg 120 cgtgggcagagctgcctttg ttttcactgc tcaggtgtaa acagtgtcct tttcgtcatt 180 tatttagtgccatgtttttt accttttttt tttttttctt ttttccctga gacagggttt 240 tgctctgtcacccaggctgg agtgcagtgg tgtgatctca tctcactgca acctccacct 300 cccaggcttaagctgtcctc ctgcctcagc cttccaagta gctgggacta caggtgtgga 360 ccatcatgtctgggtcattt tttgtatttt ttgacatgtt acccaggctg gtcttaaact 420 cctgggctcaagtgatcctt ccgcctcggc ctcccaaagt gctgggatta caggtgtgag 480 ctaccgcgcccggccatttt gcacattttt gtgctttatg ttggtgattt tggtttttga 540 aatggccccgtgatgtagtg cagtctagtg ttcctgggtg caagaaggtt atgacgtgca 600 gagaaaatccctgtgctaga gaagcacaag ctaaggcatg agttaatgtg ctgttggcca 660 cgagttcaatgagaatgaat caatagtata tgttaaatag agtcttttaa gcacaaacag 720 acataaaataggtttatgta ttgattggtt gacaaaaatg tcatgcccaa agactggcag 780 gaacctaaccctgtgtttcc cctgtgagca gtggttcagc attgactaat tcactgttct 840 tcatgacttgaaagagcata actgcagtat atgatgacga gtaaccgtaa ttcaaagtag 900 ttgatatgtgctcagagatg ctgctgagaa agtctgggaa ctactgtttc tctggaactt 960 tcctttgtgtggtagagggc ggagggccta tgagttcctc tctgggagtc ccttgtgctg 1020 ctgactgtgtcccttccttc catgtgggta ggttggagcc ataatcatca cccccactcg 1080 agagctggccattcaaatag acgaggtcct gtcgcatttc acgaagcact tccccgagtt 1140 caggtgaattggatgcagtg tccctgttag tcatgggctg ttttgtcgaa cttaatcaaa 1200 ggctgttttcttgttgtagc cagattcttt ggatcggagg caggaatcct ggagaagatg 1260 ttgagaggtttaagcaacaa gggtgagttt gctcgtgtct gcttgtttct ttgcttgctt 1320 ttggcattgaacctaagaat cgatgtgtga tctcagccta actgtgggtc cccctttcag 1380 cagcctcctccccctgcaaa aaggtgtttt ctccaggcca gcttcccatt gtgctttgct 1440 ggctccttcccataaggccc tgatgagaac ttcacaaact caggttctta cagaggtcaa 1500 gagaacgacgtgaagataaa tgagtggtca ggggctgggg gatggtgagg aactggccag 1560 cagctgccttgaatactcag ctccagctgg tagttgcctg ggaggactcg gctcagtagt 1620 ggcaggttttgaatttttta aaaagaagat ggaattccag atttgtgtgt gtaatctcct 1680 gattttttaactgttggcgt gtaatccaca tttgaaaaac atatcttgta ggtgagccag 1740 tctgagggagccacatctgg ccctgagtct gccagtttgt gatctttaca taggattctc 1800 taattctgtcaacatttcca ttttacaagt gagttacctg ggacctacaa tggttgagta 1860 acttgcgtaagtcagaggtt ggtaagcttt ttttttataa aggagcagat ggtaaatgtt 1920 ttcggctttgtgaaccatat ggtctccatc gcacttactc aactgctgtt gtagtgtgaa 1980 ggcagcgatagacaacatgt gaataaatgg gcgtggtttt ctgccaataa aatacttggc 2040 aaggacaggcagcagcccag gtttggtccg ggagctggag tttgccaacc ttgcctaggt 2100 catagcccagaacccagatc ccgtctcacc acagccctca gtctttctac cctgcagcac 2160 tcttctactctcatgctctt cgttctcttc actagggctt aaatgacttt ggggggatat 2220 aaaacttggtattttggtaa tcatcaacca cattcccaag atccagagag tttggtaaat 2280 gttaactttctttccaaagt gggaacatca ttgtggccac tccaggccgc ttggaggaca 2340 tgttccggaggaaggccgaa ggcttggatc tggccagctg tgtgcgatcc ctggatgtcc 2400 tggtgttggatgaggcagac agacttctgg acatggggtt tgaggcaagc atcaacacca 2460 ttctggagtttttgccaaag cagaggagaa caggcctttt ctctgccact cagacgcagg 2520 aagtggagaacctggtgaga gcgggcctcc ggaaccctgt ccgggtctca gtgaaggaga 2580 agggcgtggcagccagcagt gcccagaaga ccccctcccg cctggaaaac tactacatgg 2640 tatgcaaggcagatgagaaa tttaatcagc tggtccattt tcttcgcaat cataagcagg 2700 agaaacacctggtcttcttc aggcacctgt gcctgtgtgg aatactatgg gaaggctctg 2760 gaagtgctggtgaagggcgt gaagattatg tgcattcacg gaaagatgaa atataaacgc 2820 aataagatcttcatggagtt ccgcaaattg caaaggtggg attttagtgt gcactgatgt 2880 gatggcccggggaattgata ttcctgaagt caactgggtt ttgcagtatg accctcccag 2940 caatgcaagtgccttcgtgc atcgctgcgg tcgcacagct cgcattggcc acgggggcag 3000 cgctctggtgttcctcctgc ccatggaaga gtcatacatc aatttccttg caattaacca 3060 aaaatgccccctgcaggaga tgaagcccca gagaaacaca gcggaccttc tgccaaaact 3120 caagtccatggccctggctg acagagctgt gtttgaaaag ggcatgaaag cttttgtgtc 3180 atatgtccaagcttatgcaa agcatgaatg caacctgatt ttcagattaa aggatcttga 3240 ttttgccagccttgctcgag gttttgccct gctgaggatg cccaagatgc cagaattgag 3300 aggaaagcagtttccagatt ttgtgcccgt ggacgttaat accgacacga ttccatttaa 3360 agataaaatcagagaaaagc agaggcagaa actcctggag caacaaagaa gagagaaaac 3420 agaaaatgaagggagaagaa aattcataaa aaataaagct tggtcaaagc agaaggccaa 3480 aaaagaaaagaagaaaaaaa tgaatgagaa aaggaaaagg gaagagggtt ctgatattga 3540 agatgaggacatggaagaac ttcttaatga cacaagactc ttgaaaaaac ttaagaaagg 3600 caaaataactgaagaagaat ttgagaaggg cttgttgaca actggcaaaa gaacaatcaa 3660 gacagtggatttagggatct cagatttgga agatgactgc tgattccagt gccacagatg 3720 aacccacaaggacatagctg ttccctaact tggtggatgg ctccagtttg cttttaacga 3780 aaatcacaacttcaggagac atctgaaaag aatgatgtct ctgaaagctg tcctttcaga 3840 tgagggagaaatgaaggatt tcacacttca gaatatttta ctaaaaacat tccagtcttg 3900 gccgggtgcggtggctcctg cctataatcc cagcactttg ggaggctgag gtgggaggat 3960 cacttgagcccaggagttca aggctgcagt gaataatggt tgcaccattg cactctagct 4020 tgggggacagcgtgagaccc tgtctctata ttaaatttta aaatataata aaagagaaag 4080 aaaatgcctaattacagttc ttggtgaata aatgatgtaa atgcccatat tttactacct 4140 gctcacgtatagcgttagta ttgtaatcag atagtttatg taccaaactt gtaagttatg 4200 ccagttgggaccatttatct aacaggaagc aggcccaggc aggttccttt gcaacaaagg 4260 atggttcaatgttgcataca gttgagttgc catgagagac aattatggcc atccccaaat 4320 aaggaaaacttgattgcact tagataaaaa ttgcgatttt tcaaattttg aggtcaaaac 4380 ctgtatactctacaactaga ggtaagctta agctaattat gttctatttg agtcttccct 4440 tctgctgactaaatcaccat ttaaaatatt tgtaaacctt ggttgtaaag ttggacacct 4500 aagaattcaaaatagcaatt gtcccacaaa gccccatccc ttccagtatt gtgttctttt 4560 aggaaagtattcaattatct ctcagctgat cctaagattc cttctatatc ttgtcccttt 4620 catttgtttagatatttact atacactttg a 4651 <210> SEQ ID NO 141 <211> LENGTH: 147 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 141 actggcagtgcctggctcaa gaggtactca aatgtatttt aatcatttaa atgattgact 60 gcaaggtttccagctttgag atgaatagga taatgatgtc atcatgaaca cagaagtaaa 120 tgcgtgtttggcaataaaag agatggg 147 <210> SEQ ID NO 142 <211> LENGTH: 417 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure<222> LOCATION: (4) <223> OTHER INFORMATION: a, c, g or t <221>NAME/KEY: unsure <222> LOCATION: (56) <223> OTHER INFORMATION: a, c, gor t <400> SEQUENCE: 142 gaantaaaat caattagcca gatttgtaac tttaaagctaccacagtgag attttnctct 60 taaaaacatg catgttttta tattgtattc tttaatatttatattgtaca gttacatgtg 120 acacatatta catatgacta atttatatgt agcaacagggattaaagaag acctctttta 180 attgtaccac ttgttataat gctattattt acccagattcaaatgtaact acttacaatt 240 tttaaacatt tttatttctg tattctttaa aatattaaggcaatggtttt tcttgaaatt 300 atattttgcc ctatgtattc tatttttatt cacactggatttattatgat aattatatca 360 aagtgattat ttttaccctg acttttaaaa gtcaactgtttcagaagtca agttgat 417 <210> SEQ ID NO 143 <211> LENGTH: 834 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 143 ggcccctgcagggatcggcc tccagggcag gcaacagggc ctctcactga cacaggctcg 60 ggccctgcctggccagaatg cttcttccct gggagcagcc tcggtggggc cccattgagc 120 cgcagactcaagggcaggac ggagcccggt aggtcgcccc gaagcatggc gggctctcac 180 ctttgtcctgccatggagtc agagacacga gaccctggag tggggagcag gtgagtgtcc 240 tttgctcctgcctagcatcc agtaggtgcc aataaatacc tgctgactgg gtggctgaag 300 ggactgcgctggcatcccaa ggaattttgg gatccaccac cctcgagacc gtgctgggcg 360 ggaagcatgaccccacactg tgggatgggg agcctggtgg ctgccggctg gcgcgcaccc 420 caagccctttcctgggctca gctgtgagct caggacggtt tagacgcagc agcaatgatt 480 ttaagagttgccgtgttaaa cggtgatagc ctgtgacggt gctgtgtgcg gaaacatccg 540 attttattgccgctgggtgg acagcgccgt ctagacgccg ctcacccttc tcaccttcgc 600 agttgtctgtgtggaagacg ggagaggcag gggcaccacg ccactgtccc caagaccggc 660 ccccaggcactgcgcggaag aggccgtggc ctgtgaggtg gcccttccct gccctggaca 720 ggctgtggaggggctgcagg cggaggctgg gcttgggagt gcttgaagga tcactgggct 780 gtggaggggcccgcaggccg cctggggatt tggggaggga agctccgggc cttt 834 <210> SEQ ID NO 144<211> LENGTH: 982 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 144 ggcccctgca gggatcggcc tccagggcag gcaacagggc ctctcactgacacaggctcg 60 ggccctgcct ggccagaatg cttcttccct gggagcagcc tcggtggggccccattgagc 120 cgcagactca agggcaggac ggagcccggt aggtcgcccc gaagcatggcgggctctcac 180 ctttgtcctg ccatggagtc agagacacga gaccctggag tggggagcaggtgagtgtcc 240 tttgctcctg cctagcatcc agtaggtgcc aataaatacc tgctgactgggtggctgaag 300 ggactgcgct ggcatcccaa ggaattttgg gatccaccac cctcgagaccgtgctgggcg 360 ggaagcatga ccccacactg tgggatgggg agcctggtgg ctgccggctggcgcgcaccc 420 caagcccttt cctgggctca gctgtgagct caggacggtt tagacgcagcagcaatgatt 480 ttaagagttg ccgtgttaaa cggtgatagc ctgtgacggt gctgtgtgcggaaacatccg 540 attttattgc cgctgggtgg acagcgccgt ctagacgccg ctcacccttctcaccttcgc 600 agttgtctgt gtggaagacg ggagaggcag gggcaccacg ccactgtccccaagaccggc 660 ccccaggcac tgcgcggaag aggccgtggc ctgtgaggtg gcccttccctgccctggaca 720 ggctgtggag gggctgcagg cggaggctgg gcttgggagt gcttgaaggatcactgggct 780 gtggaggggc ccgcaggccg cctggggacc tggggaggga agctccgggcctttctctgg 840 ctctgcagac gcacggcctg ggttcaaatg ccagctccac cacttcctggctgtgtggcc 900 ttggacatgt gtcttaccct ctctgagcct taaatttcct tctctggaaaatcaatgctt 960 catcaaataa aaagtgatca tg 982 <210> SEQ ID NO 145 <211>LENGTH: 601 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:145 aacagccttt ctgaccccac agtgaggaga aactgaggca accgaagcct tgaggagggt 60tctttcacca gtgtgagcat agctgacttt gggattcttt tgttgtgatc tgtggatcaa 120tcctgtccct gagctctcct gattttgtta taacagcaga gtccacttag attttaacca 180tttgatcagc agagccttgt ttttggcctg ttgtgctgca tgatttacag tgtgcatctg 240gatttgtaat tatgcctcac atagtttata ttttgattca cttcaggctt ttctgctttc 300tcctctttac ctgcctctag ggccatatgg agtggaagtt tgcatgttgt tgtaaaagca 360ttttgaaatg ttcatcagac tttcatggtt tccttacttc ccttttaggt tttgcatttt 420aaattaggtt atagaattgt gaatttacca aactatattg tggattttat tcaaacagtt 480tctgtgggta tgaaatactg gtcatgtgta tgtatcaata ttttatatgg aatcattata 540tttcttgggg ttaggctgac aaaaaacaat ctgttagctt cattcatgag aggtattcag 600 a601 <210> SEQ ID NO 146 <211> LENGTH: 247 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 146 gttgatagct taatgatttcaaagaattat tatacattga aacacaaggt cagtcagtgg 60 ctggaaatga atgcacacatttgaagtgag aggactgctg agagcttgtt gggtggttgg 120 gacctcagtc ctaggttctctctgccccga ccttctggca agggggaata gagttagctc 180 aggaaacaaa ggtaaaagcccattttccac aactaggtaa accaatgtaa gcattaagga 240 tattaaa 247 <210> SEQ IDNO 147 <211> LENGTH: 424 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 147 gggggaattc agctaaaata ttaaagttgc taattctgagtcttggcaga tggctggaag 60 tccaagtcat cttttttctg agactcttgt gcatatggatggtttttggt tgggtctgct 120 atctgaagta tctgacgtgg taggtattat gggcaaatgataatgttcgt atttgaatag 180 ttgatagctt aatgatttca aagaattatt atacattgaaacacaaggtc agtcagtggc 240 tggaaatgaa tgcacacatt tgaagtgaga ggactgctgagagcttgttg ggtggttggg 300 acctcagtcc taggttctct ctgccccgac cttctggcaaggggaataga gttagctcag 360 gaaacaaagg taaaagccca ttttccacaa ctaggtaaaccaagtaagca ttaaggatat 420 taaa 424 <210> SEQ ID NO 148 <211> LENGTH: 574<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 148ctcgtcttgg ccctccaaat tgttgggatt acaaagtgct acatgcccag ccaagagttt 60taaaagggaa ggacatgacc agatttatgt taagagttta cccagacttc tgtgcataaa 120atcaattagt ggaagaaaat aggctgtgat gtgacacata aggaagccag tatagaattg 180aataagaagt aatgaagggg cttagatgag gttaatatat ttgctaccat cagagagaat 240ggaacacaca tgagaaatat ctacaaagtg aatttggtgg aacttgggaa cagttgcata 300tggtggacca gtttacaagg ctgatgctca ggctgtggct tgggtggttt gttggatgtt 360ggattaactg agacaaggag actgaccaag tagtttccct ccttctctgg gctttacttt 420cttccatagt tccttcctcc tttcatagtc tagatttcta cttctcagaa aatttctctg 480caagacggtg gaggcaaaat gtgggaaggg atgttgacat caataggacc tgttagcctc 540tgaacatctt gccaggctgg cttcagcaga ggac 574 <210> SEQ ID NO 149 <211>LENGTH: 248 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:149 ccactttggc taatgtaaaa aaaaaatgtg aggtgttctt aaaccaaaac ttctcaaatc 60ctaacttaac gtaatctacc tgaaatcaat gttatggtta ctggcattga caagactgga 120agagaaataa cttttcgtaa cagctcatct tcacatagct aatgataggt accattgttt 180tttgggcaaa gtgatggggg aggatattaa ctatttcaaa ggttgttcaa aatgactaaa 240atataaca 248 <210> SEQ ID NO 150 <211> LENGTH: 109 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 150 tttagaaagg tattcttgagcactatatga aagtaacata tttactacca attgttcttg 60 tttgaaggaa aattcctgtcttttcagtta aaaagataag ttttgtcta 109 <210> SEQ ID NO 151 <211> LENGTH:944 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 151atttgacttt attcattgct gtgcaaacct ttttttgcag catatactcg ggcactacat 60tcattcatca attcctgtga tgtccctggg ggtaattcaa ctcttcgagt cgcaattcat 120aattttgctt ctgcacacag gcggactttg aaaaatctat aataagaatc tgaaattaac 180tggtagtatt ttggctttta cttaaaatta tccctgagag agtatttaag aaaagctgtt 240caagttataa aatatataat ctgggaagaa atactgtctc atataataat tagattgtaa 300tcattgtttt aatctctgtc tgggaaccaa gattgaaagc tgacttactt ctctcttctg 360tcttgtgaac catacggagc ctattatttt aaaatatgat cagacaagta aggcttctct 420tactttgctc tgctctgatc agaagagctc atgtgaagtc tttgagattc tcttatttat 480catctttcta aaactgtgtt tttgagcttg acagtactga aaatgtctgg atgaagcaga 540aaagaaagtg atgaaatgtg tttctgagca tcagagacca tctatattgc cattaccttt 600tctagttgta tataagaatt caagattaga agaatttaga tttgttgcac atttttttcc 660tcagcatttt ttcctcttgt tttttaaaat gtattgcctc tttccccatt cagtgacact 720ggacatagga atttttaatt gtgtaatttt ctgttgcaaa aagggtaaat aaatcctttg 780tcttttgaat atcttctatg tgaaataatt gtgagtgaca tttgaaaaag tgaatctgaa 840gtcaaagtga gtaaattcct tatttcccta ttttttaaaa acctggtata tgacatggct 900acagggcaaa tgaaataaaa attgccatag ttggtatgaa aaaa 944 <210> SEQ ID NO 152<211> LENGTH: 3897 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 152 ataaggctac ggatgggcgg gacggagcag cccaccgcaa agtggcggtttacttgaggc 60 ggttacctta gtactccgag tagactgagt ctgtggcgag ctgcgggccgattcctggcc 120 agtgccatct cagccggagc aggcctcggg gcctcagaag caggcttttatctggcccga 180 ggctcccagc cgttcagcgc gtcttcccat aacctatacc gattattgggactctcggct 240 gcagacacag gagtcacaga tgctgggaag tatggcccga aagaaacctcgaaatacctc 300 aaggttgccc ctggctttaa accccctgaa gagcaaggac gtgttggcagtgctggctga 360 gaggaacgag gctatagtac cagttggggc atgggtggaa cctgcctcaccaggtagttc 420 ggaaatccca gcatatacat cagcatattt aattgaagaa gaactaaaggaacagctaag 480 aaaaaaacaa gaagctttga aacattttca gaaacaagtt aaataccgagtaaatcaaca 540 aattaggttg agaaaaaagc aacagcttca gaagtcttat gaaagagcacaaaaagaagg 600 ctccatagcc atgcagtctt cagcaacaca cttaacttcc aaaaggacaagtgtttttcc 660 aaacaatttg aatgttgcta ttggaagttc taggttacct ccttccctgatgcctgggga 720 tggaatagag gatgaagaga atcagaacga attattccaa caacaagcccaggctcttag 780 tgaaactatg aaacaggcac gtcaccggct agcatccttt aaaaccgtgattaaaaaaaa 840 gggatcagtg tttccagatg atggaaggaa aagctttctt accagagaggaagtgctttc 900 caggaaacca gcatccactg ggataaatac aggaataaga ggagagttgcccattaaggt 960 ccatcaaggt cttttagctg ctgtacctta ccagaattat atggaaaatcaggaacttga 1020 ctatgaggaa cctgactatg aggaatcttc atctcttgta actgatgagaaagggaaaga 1080 agatttgttt gggagaggcc agcaggacca gcaggctatc cattctgaagataagaacaa 1140 acctttcagc agagttcaga aagtaaaatt caaaaatcca ttatttgttctgatggaaga 1200 ggaagaacaa aagcagttac attttgaggg ccttcaggat attctgccagaagcccagga 1260 ttattttcta gaagcccaag gtgatttgct ggaaacccag ggtgatttgacaggaatcca 1320 gagtgttaag ccagataccc aggctgttga aatgaaggtt caggttactgagccagaagg 1380 ccaggccatt gagccagaag gccagcctat taagacagaa actcagggtattatgctgaa 1440 agcccagagt attgagctag aagaagggag tattgtgttg aaaacccaggattttctacc 1500 cacaaatcag gctcttctaa cgaaaaacca ggatgtttta ctcaaagaccactgtgttct 1560 ccctaaagac cagagtattc tactcaaata tcaggaccag gacttcctacccagagacca 1620 gcatgttctc cacaaagacc aagatattct gccaaaatat caggaccagaattttctacc 1680 taaggaccag aattttttat ctagagacca gcatgttctc cccaaagaccaagatattct 1740 gccaaaatat caggaccaga attttctacc taaggaccag aattttttgtctagagacca 1800 gcatgttctc cccaaagacc agaatattct acctaaatat caaggccaggattttctacc 1860 taaagaccag gactttttat ctagagacca gcatgttctc cccaaagactggaatattct 1920 acccaaatgt caggaccagg attttctacc cagagaccaa ggtgttcttcccaaagacca 1980 aaatattcta cccatatgtc aggaccagga ttttctaccc agagaccaaggttatcttcc 2040 taaagaccaa aatattctac ccatatgtca ggaccgggat tttctacccagagacctgca 2100 tgttctctcc aacgaccaga atattctacc caaatgtcag gaccaagattttctaccaaa 2160 atatcagaaa gtacacttta aggagccata ctctgatatg acagatgagaaagggagaga 2220 agacttttct ctggcagact atcagtgttt gcctcccaaa tcccaggaccaggatgacat 2280 caaaaatcag caacctgcat cttttatgag agaagaaaga gtgagagaggaattgcctct 2340 ggactatcat caatatgttg tacctaaaat ccaggaccaa gactcccctagagaacagaa 2400 caagcatatc aaactaccct catcttttga gaaatgggag attgcaagaggaaatactcc 2460 tggagtgcca ttggcttatg ataggtatca atcaggattg agcactgaattccaagctcc 2520 actggcattt cagtctgacg tggataaaga agaagataag aaagagcgtcaaaagcagta 2580 cctgagacat agacgacttt tcatggatat tgagagagaa caagttaaagaacaacaaag 2640 gcaaaaagaa caaaagaaga aaattgaaaa aattaagaaa aagagagagcaagaatgtta 2700 tgctgcagag cagaggatcc taagaatgaa ctttcatgaa gatccatattcaggagagaa 2760 gttgagtgag atattagccc agttacaact tcaagaaata aaaggaaccagagaaaaaca 2820 acagagagaa aaagaatacc tgagatatgt agaagcttta cgagcccaaatccaggagaa 2880 aatgcagctg tataatatta ctttacctcc actatgctgt tgtggtcctgatttttggga 2940 tgctcatcct gatacctgtg ccaacaactg tattttctat aaaaaccacagagcatatac 3000 tcgggcacta cattcattca tcaattcctg tgatgtccct gggggtaattcaactcttcg 3060 agtcgcaatt cataattttg cttctgcaca caggcggact ttgaaaaatctataataaga 3120 atctgaaatt aactggtagt attttggctt ttacttaaaa ttatccctgagagagtattt 3180 aagaaaagct gttcaagtta taaaatatat aatctgggaa gaaatactgtctcatataat 3240 aattagattg taatcattgt tttaatctct gtctgggaac caagattgaaagctgactta 3300 cttctctctt ctgtcttgtg aaccatacgg agcctattat tttaaaatatgatcagacaa 3360 gtaaggcttc tcttactttg ctctgctctg atcagaagag ctcatgtgaagtctttgaga 3420 ttctcttatt tatcatcttt ctaaaactgt gtttttgagc ttgacagtactgaaaatgtc 3480 tggatgaagc agaaaagaaa gtgatgaaat gtgtttctga gcatcagagaccatctatat 3540 tgccattacc ttttctagtt gtatataaga attcaagatt agaagaatttagatttgttg 3600 cacatttttt tcctcagcat tttttcctct tgttttttaa aatgtattgcctctttcccc 3660 attcagtgac actggacata ggaattttta attgtgtaat tttctgttgcaaaaagggta 3720 aataaatcct ttgtcttttg aatatcttct atgtgaaata attgtgagtgacatttgaaa 3780 aagtgaatct gaagtcaaag tgagtaaatt ccttatttcc ctattttttaaaaacctggt 3840 atatgacatg gctacagggc aaatgaaata aaaattgcca tagttggtatgaaaaaa 3897 <210> SEQ ID NO 153 <211> LENGTH: 542 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 153 tagaatagaa atattaaggaatctggctat ttatcctaga ttctattcca agcgcactca 60 tgacagtgat ttcaagtgaccactttaaaa gaattttcat tctgaacatt ttaatttttt 120 gtttattcat gtttttgacctgaacccttt tcttcgattt attattttat gtttggttac 180 ttccttctga aacggcttcctttcattctt ttttgatttt attctttata ttcatgtttt 240 ttccccttct ttatccaattgagcccaagt ttgcaaaaga aaagaacaga taaaaatatc 300 aaaattgttc atggggtagtttttggtaat tttgtcactt tgtgtgactg gaccttctta 360 acaggcttat gataggtatcaatcaggatt gagcactgaa ttccaagctc cactggcatt 420 tcagtctgac gtggataaagaagaagataa gaaagaggta tgtaatgata ctgcttttgg 480 atcccaatat ttctactatgatagtattaa tatatgagaa attggaaaca atttgttgtg 540 tc 542 <210> SEQ ID NO154 <211> LENGTH: 869 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 154 ggtagttttt ggtaattttg tcactttgtg tgactggacc ttctaacaggcttatgatag 60 gtatcaatca ggattgagca ctgaattcca agctccactg gcatttcagtctgacgtgga 120 taaagaagaa gataagaaag agcgtcaaaa gcagtacctg agacatagacgacttttcat 180 ggatattgag agagaacaag ttaaagaaca acaaaggcaa aaagaacaaaagaagaaaat 240 tgaaaaaatt aagaaaaaga gagagcaaga atgttatgct gcagagcagaggatcctaag 300 aatgaacttt catgaagatc catattcagg agagaagttg agtgagatattagcccagtt 360 acaacttcaa gaaataaaag gaaccagaga aaaacaacag agagaaaaagaatacctgag 420 atatgtagaa gctttacgag cccaaatcca ggagaaaatg cagctgtataatattacttt 480 acctccacta tgctgttgtg gtcctgattt ttgggatgct catcctgatacctgtgccaa 540 caactgtatt ttctataaaa accacagagc atatactcgg gcactacattcattcatcaa 600 ttcctgtgat gtccctgggg gtaattcaac tcttcgagtc gcaattcataattttgcttc 660 tgcacacagg cggactttga aaaatctata ataagaatct gaaattaactggtagtattt 720 gggcttttac ttgaaaatca tccctgagag agtattaaga aaagctgttcaagttataaa 780 atatataatc tggaaagaaa tactgtctca tataataatt agattgtaatcattgtttta 840 atctctgtct gggaaccaag attgaaagc 869 <210> SEQ ID NO 155<211> LENGTH: 1373 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 155 ggtagttttt ggtaattttg tcactttgtg tgactggacc ttctaacaggcttatgatag 60 gtatcaatca ggattgagca ctgaattcca agctccactg gcatttcagtctgacgtgga 120 taaagaagaa gataagaaag agcgtcaaaa gcagtacctg agacatagacgacttttcat 180 ggatattgag agagaacaag ttaaagaaca acaaaggcaa aaagaacaaaagaagaaaat 240 tgaaaaaatt aagaaaaaga gagagcaaga atgttatgct gcagagcagaggatcctaag 300 aatgaacttt catgaagatc catattcagg agagaagttg agtgagatattagcccagtt 360 acaacttcaa gaaataaaag gaaccagaga aaaacaacag agagaaaaagaatacctgag 420 atatgtagaa gctttacgag cccaaatcca ggagaaaatg cagctgtataatattacttt 480 acctccacta tgctgttgtg gtcctgattt ttgggatgct catcctgatacctgtgccaa 540 caactgtatt ttctataaaa accacagagc atatactcgg gcactacattcattcatcaa 600 ttcctgtgat gtccctgggg gtaattcaac tcttcgagtc gcaattcataattttgcttc 660 tgcacacagg cggactttga aaaatctata ataagaatct gaaattaactggtagtattt 720 gggcttttac ttaaaatcat ccctgagaga gtattaagaa aagctgttcaagttataaaa 780 tatataatct ggaaagaaat actgtctcat ataataatta gattgtaatcattgttttaa 840 tctctgtctg ggaaccaaga ctgaaagctg acttacttct ctcttctttcttgtgaacca 900 tacggagcct attattttaa aatatgatca gacaagtaag gcttctcttactttgctctg 960 ctctgatcag aagagctcat gtgaagtctt tgagattctc ttatttatcatctttctaaa 1020 actgtgtttt tgagcttgac agtactgaaa atgtctggat gaagcagaaaagaaagtgat 1080 gaaatgtgtt tctgagcatc agagaccatc tatattgcca ttaccttttctagttgtata 1140 taagaattca agattagaag aatttagatt tgttgcacat ttttttcctcagcatttttt 1200 cctcttgttt tttaaaatgt attgcctctt tccccattca gtgacactggacataggaat 1260 ttttaattgt gtaattttct gttgcaaaaa gggtaaataa atcctttgtcttttgaatat 1320 cttctatgtg aaataaaaaa aaaaaaaaaa aaaaaaaaaa aaaattggcggcc 1373 <210> SEQ ID NO 156 <211> LENGTH: 338 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 156 ctcctttttc aaaccgaaatctgcagtgtt tggtgcagtt tcaagaacaa agtagtgttc 60 aacgaatact tgttgaataaatgaagggaa gaattcttga aatcattctc ccttttgctt 120 aaagattgca gtgaacttacaccaaattag gcattcattg aattagctaa tattactttg 180 taaaaatata agtaccttcagtagagtcag aaactctcag gttaatagaa atgaattagt 240 ttagatttct gtcttcttaacatctaaatg aaattagctg cattgaagaa acataagttt 300 aattgagggt gcaaggtgcaggggagagtc tgggagaa 338 <210> SEQ ID NO 157 <211> LENGTH: 56 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: unsure<222> LOCATION: (45)..(46) <223> OTHER INFORMATION: a, c, g or t <400>SEQUENCE: 157 gggaggggat ggagagtgct tgatggatac agggttattt tttgnnggggcggggg 56 <210> SEQ ID NO 158 <211> LENGTH: 613 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 158 ttaacttcat tttaaggtagatgaagattt agttatctta tgttcaccct ttctccatac 60 cctccactcc caccatcactgccacatcat ctgtcctctt ccccattctc ccagtattgt 120 aatatagttt gatcagaactcttggttaag tcactagtgt ttacattact atggctgtgt 180 cactatttct ctctgctgacccaagcatga cactgattag gtttcctttc tcatataact 240 cttgtccttg gattcagtggccctcatttt tttcttttgc tttgttttct gttactgttc 300 atcacatatt ttatacagctgtcgatgtta tttattcaaa cgatgtacca gttccatttg 360 tttgtttgtt tctggagactccctctggag ctttccacct tcctggctct aatctggact 420 ggttgcttta ggcctgtggtctaaacagct gttatttcag gacttccttg actgttctcc 480 tttgtttgcc ctttttttctgatctcatgg gttttttgtg tgtacttttt ttatcttgct 540 tacaccctca ttttgctggtacttatcttt cagtggcttt tctaagaaag attatgtagg 600 aagtagattt ttt 613 <210>SEQ ID NO 159 <211> LENGTH: 6239 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 159 tttttttttt agctgggtgt agtggtgcgt gcctgtagtcccagctacgt gagaggccga 60 ggtgggatga tctcttgagc ctaggaggtt gaggctgcagtgagtggtga ttgcactact 120 gcactctagc ctaggctata gagtgagacc ctgtctcaaaaatacaaata aataaattaa 180 ataaaagaag agggctgtcc cagttcttca gaataaccaaaggtggaatg gtgagaggac 240 agtattttca tttactttct aagcattaat acctcaataattgggaaaaa aattaaaggc 300 aagaagatga aaaataagaa aaaagaggac atttaggagatcagtccagg agggtttaag 360 atccaagtgc taagcacaca aattaaacaa acaaacaaaaagtcacatag aaggaatgag 420 aatgacatgg aacctttaaa agaaaaaaag caatgtctataaattatgaa gaaaaataat 480 tttaaactca aaattatatc ccagctaggt tactaatcaaaattagggtg gaataaagat 540 atttttaaat ataaaagaac tatgggggat ggtctcacatttggaaaaac aaatcaaata 600 tacaagcagt caaaaaaatc tacttcctac ataatctttcttagaaaagc cactgaaaga 660 taagtaccag caaaatgagg gtgtaagcaa gataaaaaaagtacacacaa aaaacccatg 720 agatcagaaa aaaagggcaa acaaaggaga acagtcaaggaagtcctgaa ataacagctg 780 tttagaccac aggcctaaag caaccagtcc agattagagccaggaaggtg gaaagctcca 840 gagggaggtc tccagaaaca aacaaacaaa tggaactggtacatcgtttg aataaataac 900 atcgacagct gtataaaata tgtgatgaac agtaacagaaaacaaagcaa aagaaaaaaa 960 tgagggccac tgaatccaag gacaagagtt atatgagaaaggaaacctaa tcagtgtcat 1020 gcttgggtca gcagagagaa atagtgacac agccatagtaatgtaaacac tagtgactta 1080 accaagagtt ctgatcaaac tatattacaa tactgggagaatggggaaga ggacagatga 1140 tgtggcagtg atggtgggag tggagggtat ggagaaagggtgaacataag ataactaaat 1200 cttcatctac cttaaaatga agttaatggg cagtgtttaaaattgataag aaatagccat 1260 aagagcatat ttgaaaatat gaaggaaaaa aatagctcaagtggttgcct ctgggcaggg 1320 gattagaagg gatgggagag aaaactacag tttttgttataagcttttcc tgccatttga 1380 tttgaaactg ggtccctccc tctatatgtc aaaattacaatttgagatga gatttgggtg 1440 gggacacaga gccaaaccat atcaaactct gtgagaaaatttcaaactgc catcttagct 1500 cccagcccac ccacactggt tcttctacct ggctccttttgctgcaggga gctgcccaag 1560 gtgtccctga ctatacccct gatgatgata gcaatgagcagacgggggct ctgcgggagt 1620 ggtgcagcca tagcaggatg ctgggcattg gggttcaaagagttcaaagg cgtgaagagc 1680 tgtcccagag aggtgaccct gagagacatc atttattgagagtttactcc aagccaagca 1740 tagggctaca tgggcttcat atgtgttgtt tccgatccttacaacaacac atcaaacttt 1800 cacgcttcga tgaagcaagt agccatgttg aggaggttcacgtggcaaaa aagcgagaga 1860 agcttctggc caaagccagc aagttgctga atctcaccaacaacactgtg atcttggaag 1920 cagattcttc cccaattgag gatcagataa agcccagtcctggccaacac cttaagtgta 1980 gtctgaagcc gtggactgag ctaaaccaca ctgcatattacagaaactgt gagataataa 2040 atacatgttg tttcaagctg ctaagtttct gcccgctgcccaggccatgc cgccccatct 2100 gcgcgcggag ccgcggctgc cgggcctccg gggctgagccgggagcgccg ggaggaggag 2160 gcgccggcgg cggagcagga gcgggagccg cggcggcgggcagcgcggga cccagtacta 2220 tggctgtgta ctgctatgcg ctcaatagcc tggtgatcatgaatagcgcc aacgagatga 2280 agagcggcgg cggcccgggg cccagtggca gcgagacgcccccgcccccg aggagggcag 2340 tgctgagccc cggcagcgtt ttcagccccg ggagaggcgcctctttcctc ttccccccag 2400 ccgagtcgct gtcccccgag gagccccgga gccccgggggctggcggagc ggccggcgca 2460 ggctgaatag tagcagcggc agtggcagcg gcagcagcggcagtagcgtg agcagcccaa 2520 gttgggctgg tcgcctgcga ggggaccggc agcaggtggtggcagccggt accctctccc 2580 cgccagggcc ggaggaggcc aagaggaagc tgcggatcttgcagcgcgag ttgcagaacg 2640 tgcaggtgaa ccagaaagtg ggcatgtttg aggcgcacatccaggcacag agctccgcca 2700 ttcaagcgcc ccgcagcccg cgtttgggca gggctcgctcgccctccccg tgccccttcc 2760 gcagcagcag tcagccccct ggaagggtcc tggttcagggcgcccggagc gaggaacgga 2820 ggacaaagtc ctggggggag caatgtccag agacttcaggaaccgactcc gggaggaaag 2880 gagggcccag cctatgctcc tcgcaggtga agaaaggaatgccacctctt cccggccggg 2940 ctgcccctac aggatcagag gctcagggtc catccgcttttgtaaggatg gagaagggta 3000 tccctgccag tccccgctgt ggctcaccca cagctatggaaattgacaaa aggggctctc 3060 ctaccccggg aactcggagc tgcctagctc cctcattggggctgttcgga gctagcttaa 3120 cgatggccac ggaagtggca gcgagagtta catccactgggccacaccgt ccacaggatc 3180 ttgccctcac tgagccgtct gggagagccc gtgagcttgaggacctgcag cccccagagg 3240 ccctggtgga gaggcagggg cagtttctgg gcagtgagacaagcccagcc ccagaaaggg 3300 gcgggccccg cgatggagaa ccccctggga agatggggaaaggatatctg ccctgtggca 3360 tgccgggctc tggggagcct gaagtgggca aaaggccagaggagacgact gtgagcgtgc 3420 aaagcgcaga gtcctctgat tccctgagct ggtccaggctgcccagggcc ctggcctccg 3480 taggccctga ggaggcccga agtggggccc ccgtgggcggggggcgttgg cagctctccg 3540 acagagtgga gggagggtcc ccaacgctgg gcttgcttgggggcagcccc tcagcacagc 3600 cggggaccgg gaatgtggag gcgggaattc cttctggcagaatgctggag cctttgccct 3660 gttgggacgc tgcgaaagat ctgaaagaac ctcagtgccctcctggggac agggtgggtg 3720 tgcagcctgg gaactccagg gtttggcagg gcaccatggagaaagccggt ttggcttgga 3780 cgcgtggcac aggggtgcaa tcagagggga cttgggaaagccagcggcag gacagtgatg 3840 ccctcccaag tccggagctg ctaccccaag atccggacaagcctttcctg aggaaggcct 3900 gcagccccag caacatacct gctgtcatca ttacagacatgggcacccag gaggatgggg 3960 ccttggagga gacgcaggga agccctcggg gcaacctgcccctgaggaaa ctgtcctctt 4020 cctcggcctc ctccacgggc ttctcctcat cctacgaagactcagaggag gacatctcca 4080 gtgaccctga gcgcaccctg gaccccaact cagccttcctgcataccctg gaccagcaga 4140 aacctagagt gaaatacaga accatctgga aggtgaagaacaaagaaagg gaatcaagcc 4200 ctgggaatgc aagtttgttg cttattcctg tgactgctgccacagggata agggtgctag 4260 gcttgggttt aggagacttg ggtgaaattc cagtctacacatggctagca agctctctga 4320 aaaatgggga gagtaagtgt gacctcatgg agtggtactgctatactgtg aagcacccag 4380 gcagcctgga gctgcacggc ctgcgcatga gtccaacaggtacaagctgt tgtgggctta 4440 taatgtcagc acctaagcag gaattgaatg caatcgagttgagttacctg cctccagctc 4500 ccatagttgt ggtgaggaaa agtggcttca gtgctcagcagagtgcttgg gactgcatca 4560 agcccagttc gcctatcagg gacagggtag ctctcctgtgccctatgggc ttcaaggcaa 4620 aagggcttta tgaatcctgc ctctggcaca gcccagagtccagtggtatc cggcagaagc 4680 agtgctgtgc agctctgagc tgggccctta aggggaagagggagtacctc cagcaatatt 4740 cagggtggat gtgggttcca gggcttctga tcctggggctgggcttgtct gagattcaca 4800 ggagctcctt gcaagttcag cctgctggag gtgtacacacagaagcagca gcccctggag 4860 cccctggaca ccagggagcc atgtccgtca catatgatgccttgagggag aaacagcagc 4920 tcagcaaagt aggtgacttg cccgcgttga catggccaggtcctcttatc agccagatgc 4980 cgggagtatt ggattcctgc cggctgtgca gcctgggggacatagagaag agcaaatcat 5040 ggaggaagat aaaaaacatg gtgcactggt ctcccttcgtcatgtccttc aagaagaagt 5100 acccctggat ccagctggca ggacacgcag ggagtttcaaggcagctgcc aatggcagga 5160 tcctgaagaa gcactgtgag tcagagcagc gctgcctggaccggctgatg gtggatgtgc 5220 tgaggccctt cgtacctgcc taccatgggg atgtggtgaaggacggggag cgctacaacc 5280 agatggacga cctgctggcc gacttcgact cgccctgtgtgatggactgc aagatgggaa 5340 tcagacagca gcaggacttc gcaggtgacc acatggagaataatccaagt ggtgtccact 5400 cagacctggc caaaaaagca ggggagtgtg gggaggggctgagcctcacc ttcctgtggg 5460 catcccgccc caccatccag ctggcacccc ctgtggacatctccccccag cctctttcct 5520 ctcctgggca gacctacctg gaggaggagc tcacgaaggcccggaagaag cccagcctgc 5580 ggaaggacat gtaccagaag atgatcgagg tggaccccgaggcccccacc gaggaggaaa 5640 aagcacagcg ggctgtgacc aagccacggt acatgcagtggcgggagacc atcagctcca 5700 cggccaccct ggggttcagg atcgagggaa tcaagctaagaggctctgcc tggggtgcac 5760 tgcccacagc ccccggctct cggcccctcc tgcacccagggctgctccct cagccccagg 5820 tcctgccagt cctgtcgaag gcagccacaa aagaagacggcaccgtgaac cgggacttca 5880 agaagaccaa aacgagggag caggtcaccg aggccttcagagagttcact aaaggaaacc 5940 ataacatcct gatcgcctat cgggaccggc tgaaggccattcgaaccact ctagaagttt 6000 ctcccttctt caagtgccac gaggtcattg gcagctccctcctcttcatc cacgacaaga 6060 aggaacaggc caaagtgtgg atgatcgact ttgggaaaaccacgcccctg cctgagggcc 6120 agaccctgca gcatgacgtc ccctggcagg aggggaaccgggaggatggc tacctctcgg 6180 ggctcaataa cctcgtcgac atcctgaccg agatgtcccaggatgcccca ctcgcctga 6239 <210> SEQ ID NO 160 <211> LENGTH: 15 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 160 Met Asn AsnSer Gly Ala Asp Leu His Leu Ser Thr Gly Thr Ile 1 5 10 15 <210> SEQ IDNO 161 <211> LENGTH: 37 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<400> SEQUENCE: 161 Met Asn Tyr Lys Leu Ser Glu Ile Ile Leu Ser Ser LysLeu Ile Thr 1 5 10 15 Asp Val Ser Glu Ile Thr Gln Ile Met Phe Pro PheGln Phe Lys Ser 20 25 30 Arg Pro Phe Pro Leu 35 <210> SEQ ID NO 162<211> LENGTH: 94 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 162 Met Gly Gln Glu Ala Gly Val Trp Gln Val Ser Phe Cys PheLys Lys 1 5 10 15 Gly Lys Gln Lys Glu Cys Gln Lys Phe Asp Phe Asn PheLeu Ala Glu 20 25 30 Ala Phe Leu Pro Phe Ser Cys Pro Phe Phe Phe Pro LeuPro Ser Phe 35 40 45 Pro Pro Ser Val Leu Ser Ser Phe Leu Phe Pro Leu LeuIle Pro Phe 50 55 60 His Arg Thr Phe Cys Ala Gln Lys Met Thr Ala Ser CysHis Ala Pro 65 70 75 80 Leu Cys Glu Ser Ser Cys Ser Leu His Cys Gln LeuHis Phe 85 90 <210> SEQ ID NO 163 <211> LENGTH: 53 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <400> SEQUENCE: 163 Met Thr Leu Asn Glu His AlaAla Phe Lys His Leu Phe Asn Glu Ala 1 5 10 15 His Leu Ala Pro Pro LeuIle His Leu Thr Leu Ser Gly His Ser Thr 20 25 30 Cys Phe Arg Glu His ArgVal Gly Gly Thr Val Pro Asp Thr Gly Asp 35 40 45 Asn Lys Glu Lys Gln 50<210> SEQ ID NO 164 <211> LENGTH: 31 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 164 Met Leu Ile Cys Phe Tyr Pro Asp Thr TyrAsn Gln Val Glu Leu Gly 1 5 10 15 Ile Leu Phe Ser Leu Arg Val Gly GluHis Arg Ile Thr Leu Tyr 20 25 30 <210> SEQ ID NO 165 <211> LENGTH: 36<212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 165 Met IleThr Lys Ile Ile Asn Tyr Leu Gln Ile Ile Phe Thr Gly Ile 1 5 10 15 ValArg Pro Ile Arg Lys Asn Tyr Lys Thr Leu Trp Asp Gly Tyr Lys 20 25 30 ArgArg Phe Glu 35 <210> SEQ ID NO 166 <211> LENGTH: 19 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 166 Met Phe Leu Asn Cys ThrMet Asn Tyr Lys Asn Leu Leu Ala Arg Ser 1 5 10 15 Val Leu Phe <210> SEQID NO 167 <211> LENGTH: 22 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<400> SEQUENCE: 167 Met Lys Cys Phe Ser Phe Cys Leu Asn Thr Thr Ser PheThr Val Val 1 5 10 15 Lys Val Asn Tyr Phe Pro 20 <210> SEQ ID NO 168<211> LENGTH: 68 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 168 Met Arg Leu Phe Ala Ile Val Gly Cys Trp Lys Phe Gly TyrSer Lys 1 5 10 15 Trp Tyr Ile Arg Leu Leu Phe Ala Cys Ala Pro Glu ValPhe Val Pro 20 25 30 Ala Ser Arg Ser Ala Val Ser Thr Pro Leu Ser Gln ProVal Gly Ser 35 40 45 Thr Cys Glu Lys Leu Ser Ile Pro Gly Leu Ser Gly ArgPhe Leu Thr 50 55 60 Ser Leu Met Phe 65 <210> SEQ ID NO 169 <211>LENGTH: 105 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:169 Phe Leu Leu Arg Gln Asp Leu Thr Leu Ser Pro Lys Leu Glu Cys Ser 1 510 15 Gly Ala Ile Met Ala His Cys Ser Leu Gly Leu Pro Gly Ser Ser Asn 2025 30 Pro Ser Thr Ser Ala Ser Arg Leu Ala Gly Thr Thr Gly Ala Tyr His 3540 45 Gln Ala Trp Leu Ile Phe Leu Ile Lys Thr Gly Val Tyr Tyr Val Ala 5055 60 Gln Ala Gly Leu Glu Leu Leu Asp Ser Ser Asn Ser Pro Thr Leu Ala 6570 75 80 Ser Gln Ser Asp Arg Ile Thr Gly Met Ser His His Ala Gln Pro Gly85 90 95 Ser Pro Leu Leu Thr Ile Thr Ile Pro 100 105 <210> SEQ ID NO 170<211> LENGTH: 35 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 170 Met Leu Thr Ile Ser Glu Lys Ile Ile Ser Tyr Ile Tyr IleLeu Val 1 5 10 15 Ser Lys Asp Ala Leu Lys Ala Leu Ser Ser Ile Val HisAsn Ile Pro 20 25 30 Gly Leu Phe 35 <210> SEQ ID NO 171 <211> LENGTH: 78<212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: UNSURE <222> LOCATION: (74) <400> SEQUENCE: 171 Met Ala LeuGly His Ile Ser Gln Trp Ser Asp Pro Gly Ser Gln Gln 1 5 10 15 Ser LeuLeu Ser Ile Arg Asp Arg Thr Met Ala Gly Thr Leu Ser Lys 20 25 30 Val ProHis Asp Pro Glu Asp Met Cys Glu Phe Cys Ile Ile Phe Pro 35 40 45 Ser IleIle Leu Arg Thr Val Arg Ala Lys Val Arg Thr Leu Thr His 50 55 60 Arg PheVal Thr Arg Arg Asn Ser Leu Xaa Thr Glu Ser Phe 65 70 75 <210> SEQ ID NO172 <211> LENGTH: 32 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 172 Met Arg Pro Gly Trp Pro Leu His Phe Leu Arg Asp Val MetAsn Ser 1 5 10 15 Arg Val Thr Lys Met Gln Thr Ala Ser Ser Arg His ArgGly Met Val 20 25 30 <210> SEQ ID NO 173 <211> LENGTH: 46 <212> TYPE:PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 173 Met Glu Lys Asp LeuArg Val Gln Ser Ser Gly Pro Ile Leu Pro Arg 1 5 10 15 Arg Leu Gly LysPhe Met Arg Val Ser Gly Arg Gly His Gly Val Leu 20 25 30 Ile Asp Leu PheSer Gln Leu Lys Ser Ser Phe Arg Leu Ser 35 40 45 <210> SEQ ID NO 174<211> LENGTH: 39 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 174 Met Val Cys Arg Cys Ser Arg Lys Leu Cys Arg Trp Tyr ValGly Asn 1 5 10 15 Trp Ile Trp Gly Asn Ala Ala Ala Cys His Ala Leu SerIle Gly Arg 20 25 30 Phe Ser Pro Leu Phe Pro Pro 35 <210> SEQ ID NO 175<211> LENGTH: 38 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 175 Met Asn Thr Thr Leu Leu Cys Leu Cys Arg Ile Leu Pro GluHis Gly 1 5 10 15 Gly Lys Ser Thr Gly Ile Val Val Arg Lys Leu Gly PheTrp Pro Glu 20 25 30 Phe Ala Pro Asp Tyr Gln 35 <210> SEQ ID NO 176<211> LENGTH: 36 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 176 Met Leu Ala Lys Ile Ser Lys Thr Ile Lys Pro Gly Ser IleGlu Leu 1 5 10 15 Pro Ser Ser Tyr His Lys Val Phe Pro His Phe Leu LeuIle Val Asn 20 25 30 Phe Leu Lys Lys 35 <210> SEQ ID NO 177 <211>LENGTH: 51 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: UNSURE <222> LOCATION: (26)..(32) <400> SEQUENCE: 177Met Phe Ser Ser Pro Ser Asp Cys Leu Leu Ile Pro His Leu Phe Phe 1 5 1015 Arg Ser Leu Phe Phe Ile His Trp Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa 20 2530 Ala Phe Lys Phe Leu Leu Phe Met Arg Gln Met Tyr Leu Arg Ser Ile 35 4045 Asp Val Ser 50 <210> SEQ ID NO 178 <211> LENGTH: 15 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 178 Met Leu Ala Asn Thr IleVal Ser Val Arg Lys Cys Arg Val Trp 1 5 10 15 <210> SEQ ID NO 179 <211>LENGTH: 57 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:179 Met Ser Ser Leu Leu Lys Ala Leu Thr Phe Trp Pro Gln Arg Met Ala 1 510 15 Leu Phe Val Pro Ile Arg Thr Arg Ile Leu Ile Phe Leu Leu Leu Gly 2025 30 Pro Gly Asn Gln Arg Thr Thr Asn Thr Phe Ala Arg His Leu Gln Pro 3540 45 Ser Arg Ser Gly Arg Pro Ser Leu Ser 50 55 <210> SEQ ID NO 180<211> LENGTH: 46 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 180 Met Arg Asn Ile Asn Ile Val Asp Tyr Ile Lys Ile Gly SerPhe Cys 1 5 10 15 Ser Ser Thr Met Ser Glu Gly Glu Lys Ala Ser His IleHis His Pro 20 25 30 Tyr Ala Pro Lys Thr Gly Met Pro Arg Ala Glu Phe ArgAla 35 40 45 <210> SEQ ID NO 181 <211> LENGTH: 47 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: UNSURE <222>LOCATION: (26)..(47) <400> SEQUENCE: 181 Met Leu Asn Met Pro Leu Thr IleGln Ile Met Tyr Tyr Leu Met Leu 1 5 10 15 Leu Ile Ile Val Leu Phe AsnLeu Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa 20 25 30 Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 35 40 45 <210> SEQ ID NO 182 <211>LENGTH: 45 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:182 Met Ser Thr Ile Arg Glu His Ile Ser Leu Tyr Ile Ile Val Thr Asn 1 510 15 Ile Leu Asn Tyr Lys Glu Lys Lys Lys Lys Asp Ala Lys Val Gln Arg 2025 30 Leu Asn Ser Gln His Pro Thr Asp Arg Glu Tyr Leu Gly 35 40 45 <210>SEQ ID NO 183 <211> LENGTH: 57 <212> TYPE: PRT <213> ORGANISM: Homosapiens <400> SEQUENCE: 183 Met Phe Cys Val Tyr Val Lys Pro Ser Pro ProVal Leu Phe Ile Gly 1 5 10 15 Gly Gly Leu Ile Ala Val Met Ala Ser IleAsn Gly Phe Leu Val Pro 20 25 30 Arg Pro Ser Val Val Leu Ser His Ser AspSer Arg Leu Asn Asn Met 35 40 45 Ala Lys Glu Glu Ser Arg Lys Leu Glu 5055 <210> SEQ ID NO 184 <211> LENGTH: 28 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 184 Met Leu Ile Phe Leu Phe Tyr Ser Ile ProIle Ser Arg Ala Gln Leu 1 5 10 15 Ile Gly Gln Pro Thr Thr Gly Ser ProCys Trp Val 20 25 <210> SEQ ID NO 185 <211> LENGTH: 27 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 185 Met Pro Thr Arg Val PheIle Thr His Tyr Tyr Ser Ile Phe Gly Val 1 5 10 15 Pro Val Pro Cys SerLeu Asn Asn Pro Gln Leu 20 25 <210> SEQ ID NO 186 <211> LENGTH: 25 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 186 Met Gln ArgGly Lys Glu Leu Ile Val Ala Leu Phe Glu Asn Tyr Leu 1 5 10 15 Arg ProSer Leu Gly His Phe Asn Ser 20 25 <210> SEQ ID NO 187 <211> LENGTH: 49<212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 187 Met LeuSer Gln Phe Leu Lys Met Glu Trp Glu Val Glu Ile Ser Gln 1 5 10 15 ValVal Ala Gly Leu Gln His Phe His Ile Leu Gly Tyr Ile Ile Thr 20 25 30 ArgCys Cys Leu Pro Ala Gly Ala Ile Thr Ala Ser Lys Ala Thr Cys 35 40 45 Phe<210> SEQ ID NO 188 <211> LENGTH: 113 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 188 Met Ala Thr Lys Gln Ser Pro Leu Phe TyrLeu Thr Gly Ser Ala Gly 1 5 10 15 Gly Ser Leu Val Leu Lys Pro Pro ProAsn His Pro Tyr Arg Val Ser 20 25 30 Leu Arg Ala Lys Met Met Pro Gln HisPro Arg Arg Pro Leu Leu Pro 35 40 45 His Gln Leu Gly Thr Lys Tyr Ser LeuLys Cys Phe Ala Cys Gln Thr 50 55 60 Thr Arg Lys Gly Asn Ala Val Ser ThrSer Ser Ile Cys Leu Cys Leu 65 70 75 80 Val Arg Arg Ala Leu Glu Glu PheArg Met Gln Val Lys Ser Met Glu 85 90 95 Gly Gly Ile Ser Phe Leu Ile CysLys Met Ser Leu Ile Lys Leu Ile 100 105 110 Thr <210> SEQ ID NO 189<211> LENGTH: 31 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 189 Met Pro Gln Thr Cys Thr Tyr Ser Lys Ser Asn Ile Leu LysIle Tyr 1 5 10 15 Gly Ile Asp Arg Asn Thr Phe Lys Ala Thr Ile His ThrAla Arg 20 25 30 <210> SEQ ID NO 190 <211> LENGTH: 38 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 190 Met Gln Phe Gln Ala LeuGly Arg Arg Val Pro Asp Cys Phe Leu Tyr 1 5 10 15 Thr Ala Ile Ile ProTyr Thr Ala Gly Ser Ser Phe Phe Asp Ile Leu 20 25 30 Cys Asn Cys Arg GlyLeu 35 <210> SEQ ID NO 191 <211> LENGTH: 78 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <400> SEQUENCE: 191 Met Lys Ile Pro Ala Leu SerTrp Val Trp Pro Ser Arg Asn Leu Leu 1 5 10 15 Ser Tyr Ile His Gly ValLeu Pro Phe Tyr Lys Leu Met Phe Cys Asn 20 25 30 His Pro Gly Tyr Phe ProArg Arg Lys Lys Lys Leu Val Glu Gln Gly 35 40 45 Glu Gly Cys Leu Lys PheGly Asn His Pro Trp Tyr Leu Asn Gln Gly 50 55 60 Lys Ala Leu Arg Ser LeuVal Leu Gly Asn Ile Leu Leu Tyr 65 70 75 <210> SEQ ID NO 192 <211>LENGTH: 34 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:192 Met Leu His Val Cys Ser Val Leu Ser Arg Gln Arg Leu Ala Pro Met 1 510 15 Lys Glu Ala Ser Glu Pro Ser Arg Arg Glu Val Phe Ser Leu Ser Asn 2025 30 Ser Gln <210> SEQ ID NO 193 <211> LENGTH: 325 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 193 Lys Val Ser Ile Leu SerThr Phe Leu Ala Pro Phe Lys His Leu Ser 1 5 10 15 Pro Gly Ile Thr AsnThr Glu Asp Asp Asp Thr Leu Ser Thr Ser Ser 20 25 30 Ala Glu Val Lys GluAsn Arg Asn Val Gly Asn Leu Ala Ala Arg Pro 35 40 45 Pro Pro Ser Gly AspArg Ala Arg Gly Gly Ala Pro Gly Ala Lys Arg 50 55 60 Lys Arg Pro Leu GluGlu Gly Asn Gly Gly His Leu Cys Lys Leu Gln 65 70 75 80 Leu Val Trp LysLys Leu Ser Trp Ser Val Ala Pro Lys Asn Ala Leu 85 90 95 Val Gln Leu HisGlu Leu Arg Pro Gly Leu Gln Tyr Arg Thr Val Ser 100 105 110 Gln Thr GlyPro Val His Ala Pro Val Phe Ala Val Ala Val Glu Val 115 120 125 Asn GlyLeu Thr Phe Glu Gly Thr Gly Pro Thr Lys Lys Lys Ala Lys 130 135 140 MetArg Ala Ala Glu Leu Ala Leu Arg Ser Phe Val Gln Phe Pro Asn 145 150 155160 Ala Cys Gln Ala His Leu Ala Met Gly Gly Gly Pro Gly Pro Gly Thr 165170 175 Asp Phe Thr Ser Asp Gln Ala Asp Phe Pro Asp Thr Leu Phe Gln Glu180 185 190 Phe Glu Pro Pro Ala Pro Arg Pro Gly Leu Ala Gly Gly Arg ProGly 195 200 205 Asp Ala Ala Leu Leu Ser Ala Ala Tyr Gly Arg Arg Arg LeuLeu Cys 210 215 220 Arg Ala Leu Asp Leu Val Gly Pro Thr Pro Ala Thr ProAla Ala Pro 225 230 235 240 Gly Glu Arg Asn Pro Val Val Leu Leu Asn ArgLeu Arg Ala Gly Leu 245 250 255 Arg Tyr Val Cys Leu Ala Glu Pro Ala GluArg Arg Ala Arg Ser Phe 260 265 270 Val Met Ala Val Ser Val Asp Gly ArgThr Phe Glu Gly Ser Gly Arg 275 280 285 Ser Lys Lys Leu Ala Arg Gly GlnAla Ala Gln Ala Ala Leu Gln Glu 290 295 300 Leu Phe Asp Ile Gln Met ProGly His Ala Pro Gly Arg Ala Arg Arg 305 310 315 320 Thr Pro Met Pro Gln325 <210> SEQ ID NO 194 <211> LENGTH: 33 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 194 Met Ala Ser Phe Leu Leu Ser Thr Pro AlaLys Arg Lys Pro His Pro 1 5 10 15 Leu Pro Pro Ala His Pro Arg Ile HisThr Phe Arg Gln Pro Ser Gly 20 25 30 Asn <210> SEQ ID NO 195 <211>LENGTH: 74 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:195 Met Ile Pro Thr Phe Val Leu Asp Ala Lys Tyr Ala Ala Leu Met Gly 1 510 15 Gln Pro Trp Gly Leu Cys Ala Ile Cys Val His Ile Cys Leu Leu Leu 2025 30 Asp Ser Val Ser Leu Arg Ser Phe Ser Thr Ala Gln His Leu Glu Arg 3540 45 Ala Ser Lys Ser Thr Ser Ser Leu His His Leu Ile Leu Ile Asn Pro 5055 60 Ala Arg Glu Gly Cys Thr Gly Arg Thr Ala 65 70 <210> SEQ ID NO 196<211> LENGTH: 97 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 196 Met Ala Asn Phe Cys Val Phe Ile Glu Thr Glu Gly Asn AlaVal Thr 1 5 10 15 Arg Arg Ala Leu Arg Lys Gln Ala Thr Ala Gly His CysSer Gly Lys 20 25 30 Pro Ala Phe Gln Pro Ala Pro Pro Gln Tyr Pro Arg ValHis Ser Glu 35 40 45 Asp Arg Arg Leu Gln Gln Pro Gln Ala Ala Gly Arg TrpGly Ala Pro 50 55 60 Asp Trp Ile Pro Pro Leu Gln Asp Thr Arg Lys Pro SerVal Ser Ser 65 70 75 80 Arg Asp Ser Arg Ile His Glu Lys Glu Val Ile LeuAsp Ser Leu Cys 85 90 95 Ile <210> SEQ ID NO 197 <211> LENGTH: 645 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 197 Ala Leu ArgPro Pro Ser Gly Phe His Ile Arg Cys Leu Gly Asp Val 1 5 10 15 Ser ProIle Ser Met Ser Pro Ile Ser Gln Ser Gln Phe Ile Pro Leu 20 25 30 Gly GluIle Leu Cys Leu Ala Ile Ser Ala Met Asn Ser Ala Arg Lys 35 40 45 Pro ValThr Gln Glu Ala Leu Met Glu His Leu Thr Thr Cys Phe Pro 50 55 60 Gly ValPro Thr Pro Ser Gln Glu Ile Leu Arg His Thr Leu Asn Thr 65 70 75 80 LeuVal Arg Glu Arg Lys Ile Tyr Pro Thr Pro Asp Gly Tyr Phe Ile 85 90 95 ValThr Pro Gln Thr Tyr Phe Ile Thr Pro Ser Leu Ile Arg Thr Asn 100 105 110Ser Lys Trp Tyr His Leu Asp Glu Arg Ile Pro Asp Arg Ser Gln Cys 115 120125 Thr Ser Pro Gln Pro Gly Thr Ile Thr Pro Ser Ala Ser Gly Cys Val 130135 140 Arg Glu Arg Thr Leu Pro Arg Asn His Cys Asp Ser Cys His Cys Cys145 150 155 160 Arg Glu Asp Val His Ser Thr His Ala Pro Thr Leu Gln ArgLys Ser 165 170 175 Ala Lys Asp Cys Lys Asp Pro Tyr Cys Pro Pro Ser LeuCys Gln Val 180 185 190 Pro Pro Thr Glu Lys Ser Lys Ser Thr Val Asn PheSer Tyr Lys Thr 195 200 205 Glu Thr Leu Ser Lys Pro Lys Asp Ser Glu LysGln Ser Lys Lys Phe 210 215 220 Gly Leu Lys Leu Phe Arg Leu Ser Phe LysLys Asp Lys Thr Lys Gln 225 230 235 240 Leu Ala Asn Phe Ser Ala Gln PhePro Pro Glu Glu Trp Pro Leu Arg 245 250 255 Asp Glu Asp Thr Pro Ala ThrIle Pro Arg Glu Val Glu Met Glu Ile 260 265 270 Ile Arg Arg Ile Asn ProAsp Leu Thr Val Glu Asn Val Met Arg His 275 280 285 Thr Ala Leu Met LysLys Leu Glu Glu Glu Lys Ala Gln Arg Ser Lys 290 295 300 Ala Gly Ser SerAla His His Ser Gly Arg Ser Lys Lys Ser Arg Thr 305 310 315 320 His ArgLys Ser His Gly Lys Ser Arg Ser His Ser Lys Thr Arg Val 325 330 335 SerLys Gly Asp Pro Ser Asp Gly Ser His Leu Asp Ile Pro Ala Glu 340 345 350Arg Glu Tyr Asp Phe Cys Asp Pro Leu Thr Arg Arg Ser Asn Lys Ala 355 360365 Lys Glu Arg Ser Arg Ser Met Asp Asn Ser Lys Gly Pro Leu Gly Ala 370375 380 Ser Ser Leu Gly Thr Pro Glu Asp Leu Ala Glu Gly Cys Ser Gln Asp385 390 395 400 Asp Gln Thr Pro Ser Gln Ser Tyr Ile Asp Asp Ser Thr LeuArg Pro 405 410 415 Ala Gln Thr Val Ser Leu Gln Arg Ala His Ile Ser SerThr Ser Tyr 420 425 430 Lys Glu Val Cys Ile Pro Glu Ile Val Ser Gly SerLys Glu Pro Ser 435 440 445 Ser Ala Cys Ser Leu Leu Glu Pro Gly Lys ProPro Glu Ser Leu Pro 450 455 460 Ser Tyr Gly Glu Leu Asn Ser Cys Pro ThrLys Thr Ala Thr Asp Asp 465 470 475 480 Tyr Phe Gln Cys Asn Thr Ser ThrTyr His Lys Ser Ser Leu Ser Leu 485 490 495 Leu Lys Ser His Pro Lys ThrPro Ala Asp Thr Leu Pro Gly Arg Cys 500 505 510 Glu Lys Leu Glu Pro SerLeu Gly Thr Ser Ala Ala Gln Ala Met Pro 515 520 525 Ala Ser Gln Arg GlnGln Glu Ser Gly Gly Asn Gln Glu Ala Ser Phe 530 535 540 Asp Tyr Tyr AsnVal Ser Asp Asp Asp Asp Ser Glu Glu Gly Ala Asn 545 550 555 560 Lys AsnThr Glu Glu Glu Lys Asn Arg Glu Asp Val Gly Thr Met Gln 565 570 575 TrpLeu Leu Glu Arg Glu Lys Glu Arg Asp Leu Gln Arg Lys Phe Glu 580 585 590Lys Asn Leu Thr Leu Leu Ala Pro Lys Glu Thr Asp Ser Ser Ser Asn 595 600605 Gln Arg Ala Thr His Ser Ala Arg Leu Asp Ser Met Asp Ser Ser Ser 610615 620 Ile Thr Val Asp Ser Gly Phe Asn Ser Pro Arg Cys Pro Ala Ala Leu625 630 635 640 Lys Ala Glu Ala Ser 645 <210> SEQ ID NO 198 <211>LENGTH: 29 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:198 Met Leu Leu Tyr Ser Thr Arg Gly Lys Lys His Gly Leu Tyr Pro Gln 1 510 15 Gln Ser Leu Gly Asn Arg Gly Ile Tyr Leu Gln Asn Gly 20 25 <210>SEQ ID NO 199 <211> LENGTH: 32 <212> TYPE: PRT <213> ORGANISM: Homosapiens <400> SEQUENCE: 199 Met Val Thr Lys Lys Asn Leu Lys Ser Asn AsnLeu Val Gly Ala His 1 5 10 15 Leu Glu Tyr Asn Ser Met Ser Ser Cys IleTyr Leu Ser His Ile Leu 20 25 30 <210> SEQ ID NO 200 <211> LENGTH: 38<212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 200 Leu AlaAsn Phe Arg Ile Phe Ser Arg Asp Arg Val Ser Pro Cys Trp 1 5 10 15 ProVal Ala Ser Gln Thr Pro Asp Leu Lys Ala Ser Ala Cys Leu Gly 20 25 30 LeuPro Lys Cys Trp Asp 35 <210> SEQ ID NO 201 <211> LENGTH: 53 <212> TYPE:PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: UNSURE<222> LOCATION: (12) <221> NAME/KEY: UNSURE <222> LOCATION: (15) <400>SEQUENCE: 201 Met Ser Phe Leu Phe Leu Asp Ile Ala Lys Trp Xaa Phe PheXaa Phe 1 5 10 15 Leu Phe Cys Tyr Cys Phe Leu Ile Tyr Tyr Lys Met LeuPhe Phe Tyr 20 25 30 Gly Gly Phe Lys His Pro Ile Pro Cys Pro Gly Phe LeuHis His Trp 35 40 45 Ile Leu Leu Ile Ile 50 <210> SEQ ID NO 202 <211>LENGTH: 59 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:202 Met Gln Leu Trp Gly Glu Tyr Ser Pro Tyr Phe Cys Arg Asn Asn Asn 1 510 15 Phe Glu Tyr Leu Cys Ala Thr Thr Val Ala Asn Thr Arg Leu Arg Cys 2025 30 Leu Leu Leu Leu Ser Gln Pro Cys Glu Val Lys Thr Leu Ser Leu Leu 3540 45 Thr Asp Glu Glu Thr Asp Ser Glu Asp Ile Lys 50 55 <210> SEQ ID NO203 <211> LENGTH: 18 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 203 Met Arg Cys Thr Gln Gln Phe Ser Ile Leu Ala Val Phe LysCys Thr 1 5 10 15 Ile Gln <210> SEQ ID NO 204 <211> LENGTH: 177 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 204 Met Asn PheLeu Lys Leu Ile Ala Val Phe Ile Val Phe Ser His Ala 1 5 10 15 Ser GluSer Pro Gln Asp Ser Thr Pro Asn Gln Leu Tyr Ile Trp Gly 20 25 30 Arg ThrLys Ala Leu Val Phe Phe Arg Ser Ser Thr Gly Asp Ser Asp 35 40 45 Ser ThrAla Arg Ile Lys Lys Leu Ile Asn Gly Asn Ser Met Pro Val 50 55 60 Ala GluGlu Leu Pro Trp Glu Met Ser His Thr Glu His Gln Ser Ser 65 70 75 80 PhePro Thr Pro Glu Ile Pro His Ser Leu Ala Pro Gly Thr Val Ala 85 90 95 IleSer Lys Pro Trp Phe Pro Ala Val Ser Gln Ile Ala Arg Val Gln 100 105 110Arg Val Asp Ile Asn Phe Cys Ser Trp Glu Asp Leu Ser Pro Ser Gly 115 120125 Lys Ala Thr Gly Lys Ser Arg Thr His Cys Thr Val Thr Ala Val Ser 130135 140 Ser Asn Ala Thr Thr His Ala Gly Ile Asn Asn Glu His Gly Trp Gly145 150 155 160 Ser Leu Glu Leu Leu Asn Cys Lys Ala His Lys Cys Leu AsnPhe Phe 165 170 175 His <210> SEQ ID NO 205 <211> LENGTH: 119 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 205 Met Thr SerMet Ala Glu Pro Gly Leu Ala Leu Tyr Leu Cys Gly His 1 5 10 15 Thr ValVal Trp Ser Ser Ser Ser Leu Met Val Thr Phe Val Arg Ile 20 25 30 Leu IleSer Val Phe Phe Leu Pro Gln Phe Ser Ser Ser Arg Leu Pro 35 40 45 His ProCys Ser Leu Phe Met Pro Ala Trp Val Val Ala Leu Asp Glu 50 55 60 Thr AlaVal Thr Val Gln Cys Val Leu Leu Phe Pro Val Ala Phe Pro 65 70 75 80 LeuGly Glu Arg Ser Ser His Glu Gln Lys Phe Ile Ser Thr Arg Trp 85 90 95 ThrLeu Ala Ile Cys Glu Thr Ala Gly Asn Gln Gly Leu Leu Ile Ala 100 105 110Thr Val Pro Gly Ala Lys Glu 115 <210> SEQ ID NO 206 <211> LENGTH: 33<212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 206 Met LeuIle Ser Lys Ile Ile Ile Gly Ile Lys Thr Gln Arg Tyr Leu 1 5 10 15 IleGlu Lys Ser His Arg Ser Pro Arg Ile Tyr Ile Tyr Leu Gly Leu 20 25 30 Ala<210> SEQ ID NO 207 <211> LENGTH: 126 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 207 Leu Pro Cys Ser Asn Phe Phe Phe Phe SerPhe Ser Leu Phe Leu Val 1 5 10 15 Phe Ile Phe Ser Ala Ile Ser Arg IlePhe Leu Leu Leu Ala Met Ser 20 25 30 Gln Ser Ile Met Ala Leu Ser Pro ArgLeu Glu Cys Asn Gly Ala Val 35 40 45 Ser Gly His Cys Asn Pro Cys Leu ProGly Ser Ser Asp Ser Pro Pro 50 55 60 Ser Ala Ser Gln Val Ala Gly Ile ThrGly Thr Cys His His Ala Arg 65 70 75 80 Leu Ile Phe Val Phe Leu Val GluMet Gly Phe His His Val Gly Gln 85 90 95 Ala Gly Leu Glu Leu Leu Thr SerGly Asp Leu Pro Thr Ser Ala Ser 100 105 110 Gln Ser Ala Gly Ile Thr GlyVal Ser His Arg Ala Arg Pro 115 120 125 <210> SEQ ID NO 208 <211>LENGTH: 88 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:208 Met Val Tyr Lys Leu Glu Trp His Ile Ala Phe Leu Arg Ile Leu Arg 1 510 15 Gln Arg Pro Gly Phe Gly Ala Lys Ile Lys Gly Trp Met Ser His Leu 2025 30 Pro Trp Tyr Gly Asn Ala Ser Val Leu Thr Ser Ala Gln Ser Asn Leu 3540 45 Lys Leu Ile Ser Pro Ser Arg Phe Phe Leu Leu Phe Leu Ala Arg Glu 5055 60 Lys Ile Thr Ser Ala Phe Phe Phe Arg Arg Val Lys Lys Lys Glu His 6570 75 80 His Ser Ile Ser Gln Asn Cys Ile 85 <210> SEQ ID NO 209 <211>LENGTH: 52 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:209 Met Ser Leu His Cys Val Thr Asn Thr Asp Leu Val Ser Lys Trp Cys 1 510 15 Arg Arg Thr Gln Ala Thr Thr Arg Asn Glu Pro Ser Leu Cys Asp Gln 2025 30 Gly Gly Pro Gly Arg Gln Thr Pro Ala His Glu Gly Arg Thr Val Val 3540 45 Ala Met Thr Ser 50 <210> SEQ ID NO 210 <211> LENGTH: 63 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 210 Met Arg LeuPro Asp Asp Ser Cys Pro Ser Cys Ser Gly Leu Pro Ala 1 5 10 15 Glu LysSer Cys Thr His Arg Ala Leu Leu Gly Phe Leu Thr Cys Gly 20 25 30 Ile HisAsp Pro Val Thr Pro Leu Ser Ser Val Met Val His Tyr Asn 35 40 45 Asn ArgSer Pro Asp His Gly Asn Tyr Phe Ser Ser Ser Thr Leu 50 55 60 <210> SEQID NO 211 <211> LENGTH: 104 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<400> SEQUENCE: 211 Met Asp Phe Glu Phe Ile Phe Phe Pro Leu Lys Lys GlyAsn Pro Leu 1 5 10 15 Ile Ala Lys Ser His Leu Gln Ile Val Lys Gln ThrSer Gln Ile Thr 20 25 30 Lys Cys Phe Leu Cys Lys Gln Lys Ile Cys Phe AlaGly Lys Gly Ile 35 40 45 Leu Leu Leu Asn Thr Gly Thr Val Ser Val Ile LeuArg Met Gly Thr 50 55 60 Val Pro Tyr Asn Leu Phe Leu Lys Tyr Leu Leu LeuLeu Gly Leu Ser 65 70 75 80 Gln Ala Pro Ile Phe Ser Val Val Met Lys LysAsn Tyr Gln Ala Thr 85 90 95 Ser Trp Val Phe Phe Ser Leu Phe 100 <210>SEQ ID NO 212 <211> LENGTH: 57 <212> TYPE: PRT <213> ORGANISM: Homosapiens <400> SEQUENCE: 212 Met Ile Glu Leu Leu Ser Pro Tyr Gln Leu ArgGlu Leu Phe Cys Ser 1 5 10 15 Leu Thr His Val Gly Arg Thr Val Arg TrpSer Glu Gln Trp Asn Leu 20 25 30 Leu Val Ala Gln Val Leu Glu Val Tyr SerAsn Gly Gly Arg Thr Gln 35 40 45 Leu Gly Ile Trp Phe Leu Leu Ser Lys 5055 <210> SEQ ID NO 213 <211> LENGTH: 31 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 213 Met Leu Glu Phe Gly Lys Cys Lys Phe CysPhe Ala Asp Glu Ile Phe 1 5 10 15 Leu Leu Asn Phe Asn Thr Leu Lys GlyIle Pro Val Phe Asn Tyr 20 25 30 <210> SEQ ID NO 214 <211> LENGTH: 37<212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: UNSURE <222> LOCATION: (29)..(32) <221> NAME/KEY: UNSURE <222>LOCATION: (36) <400> SEQUENCE: 214 Met Leu Ile Glu Val Phe Lys Gly IleTyr Lys Leu Asn Thr Leu His 1 5 10 15 Asn Tyr Gln Leu Asn Lys Cys PheTyr His Met Gln Xaa Xaa Xaa Xaa 20 25 30 Phe Phe Leu Xaa Arg 35 <210>SEQ ID NO 215 <211> LENGTH: 131 <212> TYPE: PRT <213> ORGANISM: Homosapiens <400> SEQUENCE: 215 Met Gly Gln Lys Ile Ser Arg Gln Pro Tyr SerGly Ser Trp Ser Leu 1 5 10 15 Phe Ser Cys Ser Asp Pro Gln Lys Ala SerLys Ser Leu Asn Leu Glu 20 25 30 Thr Arg Gln Phe Phe Leu Ile Ser Cys LeuLys Ala Val Gln Ser Ser 35 40 45 Val Asn Lys Pro Leu His Ala Gly Leu IleAsn Ala Gly Pro Leu Arg 50 55 60 Ala Met Thr Gln Glu His Gly Leu Gly SerThr Leu Lys Ser Arg Asn 65 70 75 80 His Ser Thr Asp Asn Gly Asn Phe ValGly Gly Asn Arg Leu Leu Glu 85 90 95 Leu Asn Ala Phe Val Arg Phe Leu AspLeu Gln Ile Ser Leu Cys Gly 100 105 110 Pro Ala Leu Gly Gly Lys Ala GlyIle His Asn Asn Leu Ile Asn Leu 115 120 125 Thr Gln Thr 130 <210> SEQ IDNO 216 <211> LENGTH: 57 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<400> SEQUENCE: 216 Met Glu Phe Arg Cys Gln Leu Ile Pro Arg Leu Ile LeuSer Tyr Ile 1 5 10 15 Lys Val Asn Asp Ile Leu His Glu Ile Met Leu ValGlu Pro Thr Arg 20 25 30 Leu Leu Ala Met Leu Pro Ser Leu Ser Ser Leu AspPhe Leu Phe Lys 35 40 45 Ser Leu Tyr Arg Val Thr Val Glu His 50 55 <210>SEQ ID NO 217 <211> LENGTH: 67 <212> TYPE: PRT <213> ORGANISM: Homosapiens <400> SEQUENCE: 217 Met Cys Glu Leu Pro Leu Leu Leu Cys Asn SerIle Leu Phe Met Ile 1 5 10 15 Cys Asp Val Ile Arg Lys Phe Leu Leu MetCys Gln Asn Lys Phe Asn 20 25 30 Phe Pro Leu Arg Gln Phe Ile Thr Leu PheLys Trp Asn Ile Lys Glu 35 40 45 Glu Pro Pro Ile Cys Lys Ile Leu Thr PheLys Phe Met Leu Ile Phe 50 55 60 Leu Asn Tyr 65 <210> SEQ ID NO 218<211> LENGTH: 69 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 218 Met Ser Cys Leu Ser Tyr Gly Phe Lys Tyr Leu Gln Cys IleAla Lys 1 5 10 15 Tyr Cys Ser Cys Thr Leu Gln Leu Arg Asn Thr Val LeuGly Phe Gln 20 25 30 Gln Lys Tyr Leu Arg Ile Ser His Ser Ser Leu Lys LysAsp Ala Lys 35 40 45 Asp Val Thr Gly Ile Ile Ile Val Ala Val Ser Cys ArgIle Lys Asp 50 55 60 Arg Thr Arg Tyr Gly 65 <210> SEQ ID NO 219 <211>LENGTH: 29 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:219 Met Leu Trp Ser Leu Tyr Ile Ser Phe Lys Val Val Ala Asn Lys Arg 1 510 15 Met Pro Ile Gln Gly Ile Tyr Trp His Phe His Gly Gly 20 25 <210>SEQ ID NO 220 <211> LENGTH: 26 <212> TYPE: PRT <213> ORGANISM: Homosapiens <400> SEQUENCE: 220 Met Asn Phe Asp Cys Ala Ser Ala Ile Leu AspIle Phe Val Met Ile 1 5 10 15 Gly Asn Arg Thr Ile Lys Cys Leu Ala Leu 2025 <210> SEQ ID NO 221 <211> LENGTH: 41 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 221 Met Leu Phe Leu Asn Trp Ala Pro Ser SerAsp Phe Ala Asn Leu Lys 1 5 10 15 Ser Ile Thr Cys Leu Cys Leu Ser LysAsn Pro Ser Ile Pro Ser Ser 20 25 30 Leu Ile Ala Pro Cys Tyr Ser Pro Val35 40 <210> SEQ ID NO 222 <211> LENGTH: 45 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: UNSURE <222>LOCATION: (31)..(39) <221> NAME/KEY: UNSURE <222> LOCATION: (42)..(43)<400> SEQUENCE: 222 Met Thr Ile Trp Gln Arg Tyr Phe Ser Tyr Asn Glu LysTyr Leu Cys 1 5 10 15 Pro Ile Ser Leu Lys Ser Asp Val Glu Lys Leu TyrIle Tyr Xaa Xaa 20 25 30 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ile Leu Xaa Xaa LeuLeu 35 40 45 <210> SEQ ID NO 223 <211> LENGTH: 31 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <400> SEQUENCE: 223 Met Phe Gln Ser Val Arg GluMet Ser Leu Ser Gly Ser Ile Pro Ala 1 5 10 15 Asn Asn Glu Glu Gly MetArg Gln Ala Gln Trp His Ser Arg Leu 20 25 30 <210> SEQ ID NO 224 <211>LENGTH: 48 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:224 Phe Phe Phe Phe Phe Leu Arg Gln Ser Phe Thr Leu Ser Gln Ala Gly 1 510 15 Val Ala Trp His Asp Leu Gly Ser Leu His Pro Pro Leu Pro Gly Ser 2025 30 Ser Asp Ser Arg Ala Ser Ala Ser Gln Ser Ala Arg Ile Thr Gly Val 3540 45 <210> SEQ ID NO 225 <211> LENGTH: 30 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <400> SEQUENCE: 225 Met Tyr Gln Lys Lys Pro IleArg Leu Lys Val Leu Lys Thr Arg Tyr 1 5 10 15 Lys Tyr Ser His Arg TyrVal Ser Glu Thr Tyr Leu Phe Gln 20 25 30 <210> SEQ ID NO 226 <211>LENGTH: 44 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:226 Met Asn Gln Asn Leu His His Leu Tyr Asn Lys Arg Ser Glu Ser Ile 1 510 15 Ala Cys Leu Ala Trp His Val Gly Arg Val Ala Lys Asp Gln Cys Ser 2025 30 Leu Met Tyr Phe Phe Lys Leu Ser Asn Asn Ile His 35 40 <210> SEQ IDNO 227 <211> LENGTH: 57 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<400> SEQUENCE: 227 Met Leu Ile Ser Phe Trp Leu Leu Thr His Ala Ala PheSer Gly His 1 5 10 15 His Met Ala Leu Lys Gln Arg Ser Val Cys Ile HisSer Pro Tyr Glu 20 25 30 Ala Tyr Val Asn Ile Asn His Gly Met Phe Pro AsnIle Leu Leu Ile 35 40 45 Phe Ala Ser Gln Leu Gly Ser Leu Ile 50 55 <210>SEQ ID NO 228 <211> LENGTH: 101 <212> TYPE: PRT <213> ORGANISM: Homosapiens <220> FEATURE: <221> NAME/KEY: UNSURE <222> LOCATION: (32)..(73)<400> SEQUENCE: 228 Met Phe His Val Phe Ser Cys Ser Arg Ser Asp Leu AlaThr Pro Gly 1 5 10 15 Asp Thr Phe Gly Tyr Thr Asn Arg Val Tyr Leu GlyGln Arg Trp Xaa 20 25 30 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa 35 40 45 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa 50 55 60 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Leu Asp SerPro Arg Trp Ile 65 70 75 80 Ser Pro Leu Ser Pro Thr Met Leu Val Leu LeuThr Trp Leu Leu Ile 85 90 95 Lys Gln Cys Gln Val 100 <210> SEQ ID NO 229<211> LENGTH: 88 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 229 Met Leu His Ser Pro Gly Leu Thr Arg Gly Trp Pro Gln LysArg Val 1 5 10 15 Gly Glu Ala Gly Gln Gln Gly Leu Ala Glu Ile Ile CysArg Ala Gln 20 25 30 Glu Ala Gly Glu Arg Arg Gln Phe Gln Gly Pro Phe ValArg Gln Val 35 40 45 Pro Gly Ala Gln Pro Gly Arg Gln Glu Gly Leu Ser ProSer Pro Arg 50 55 60 Gln Glu Gly Ser Gln Ala Glu Ala Pro Pro Ser Gly ThrPro Gln Pro 65 70 75 80 Thr Pro Ala Ala Leu Gly Gln Asp 85 <210> SEQ IDNO 230 <211> LENGTH: 23 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<400> SEQUENCE: 230 Met Glu Ala Thr Gly Val Thr Phe Ser Ser Phe Val PheGlu Gln Gly 1 5 10 15 Met Ser Val Leu Ser Leu Lys 20 <210> SEQ ID NO 231<211> LENGTH: 48 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 231 Met Lys Thr Glu Asp Ile Lys Cys Ala Arg Val Arg Ser LeuSer His 1 5 10 15 Ala Lys Gly Lys Val Lys Ile Ala Phe Phe His Ile ValSer Glu Val 20 25 30 Gln Leu Leu Arg Leu Ile Asn Glu Ser Cys Ser Ile LysGly Leu Thr 35 40 45 <210> SEQ ID NO 232 <211> LENGTH: 25 <212> TYPE:PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 232 Met Arg Tyr Ile HisVal Glu Phe Cys Ser Cys Gly Leu Met Ile Phe 1 5 10 15 Thr Leu Tyr SerIle Thr Phe His Gly 20 25 <210> SEQ ID NO 233 <211> LENGTH: 55 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 233 Met Leu ProThr Pro Val Pro Thr Ile Glu Ala Leu Leu Phe Met Leu 1 5 10 15 Lys CysGln Val Leu Thr Val His Gly Ser Met Glu Thr Phe Leu Leu 20 25 30 Phe SerVal Val Leu Gly Ala Ser Leu Leu Val Asn Leu Arg Lys Ile 35 40 45 Gly AspSer Val Asn Leu Glu 50 55 <210> SEQ ID NO 234 <211> LENGTH: 148 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 234 Met Gly ArgIle Arg Pro Asp His Thr Leu Leu Phe Gln Arg Gly Pro 1 5 10 15 Val ProAla Pro Leu Thr Ser Gly Leu His Tyr Tyr Thr Thr Leu Glu 20 25 30 Glu LeuTrp Lys Ser Phe Asp Leu Cys Glu Asp Tyr Phe Lys Pro Pro 35 40 45 Phe GlyPro Tyr Pro Glu Lys Ser Gly Lys Asp Ser Leu Val Ser Met 50 55 60 Lys CysSer Leu Phe Arg Phe Cys Pro Trp Ser Lys Glu Leu Pro Phe 65 70 75 80 GlnPro Pro Glu Gly Ser Ile Ser Ser His Leu Gly Ser Gly Ala Ser 85 90 95 AspSer Glu Thr Glu Glu Thr Arg Lys Ala Leu Pro Ile Gln Ser Phe 100 105 110Ser His Glu Lys Glu Ser His Gln His Arg Gln His Ser Val Pro Val 115 120125 Ile Ser Arg Pro Gly Ser Asn Val Lys Pro Thr Leu Pro Pro Ile Pro 130135 140 Gln Gly Arg Arg 145 <210> SEQ ID NO 235 <211> LENGTH: 940 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 235 Glu Tyr ThrSer Phe Ser Ala Leu His Asn Thr Tyr Ser Lys Ile Asp 1 5 10 15 His IleVal Gly Ser Lys Ala Leu Leu Ser Lys Cys Lys Arg Thr Glu 20 25 30 Met IleThr Asn Cys Leu Ser Asp His Ser Ala Ile Lys Leu Glu Leu 35 40 45 Arg IleLys Lys Leu Thr Gln Asn Cys Ser Thr Thr Trp Lys Leu Asn 50 55 60 Asn LeuLeu Leu Asn Asp Tyr Cys Val His Asn Lys Met Lys Ala Glu 65 70 75 80 IleLys Met Phe Phe Glu Thr Asn Glu Asn Lys Asp Thr Thr Tyr Gln 85 90 95 AsnLeu Trp Asp Thr Phe Lys Ala Val Cys Arg Gly Asn Phe Ile Ala 100 105 110Leu Asn Val His Lys Arg Lys Gln Glu Arg Ser Lys Ile Asp Thr Leu 115 120125 Ile Ser Gln Leu Lys Glu Leu Glu Lys Gln Glu Gln Thr His Ser Lys 130135 140 Ala Ser Arg Arg Gln Glu Ile Thr Lys Ile Arg Ala Glu Val Lys Glu145 150 155 160 Ile Glu Thr Gln Lys Thr Phe Lys Arg Ile Asn Glu Ser ArgAsn Trp 165 170 175 Phe Phe Glu Arg Ile Ser Lys Ile Asp Arg Pro Leu AlaArg Leu Ile 180 185 190 Lys Lys Lys Arg Glu Lys Asn Gln Ile Asp Ala IleAsn Thr His Asp 195 200 205 Lys Gly Asp Ile Thr Thr Asp Pro Thr Glu IleGln Thr Thr Ile Arg 210 215 220 Glu Tyr Tyr Lys His Phe Tyr Ala Asn LysLeu Glu Asn Leu Glu Glu 225 230 235 240 Met Asp Lys Phe Leu Asp Thr TyrThr Leu Pro Arg Leu Asn Gln Glu 245 250 255 Glu Ala Glu Ser Leu Asn ArgPro Ile Thr Asp Ser Glu Ile Ala Ala 260 265 270 Ile Ile Asn Ser Leu ProThr Lys Lys Ser Pro Gly Pro Asp Gly Phe 275 280 285 Thr Pro Lys Phe TyrGln Arg Tyr Lys Glu Glu Leu Val Pro Phe Leu 290 295 300 Leu Lys Leu PheGln Ser Ile Thr Lys Glu Gly Ile Leu Pro Asn Ser 305 310 315 320 Phe TyrGlu Ala Asn Ile Ile Leu Ile Leu Lys Pro Gly Arg Asp Thr 325 330 335 ThrLys Lys Arg Glu Phe Arg Pro Ile Ser Met Met Ile Ile Asp Ala 340 345 350Lys Ile Leu Ser Lys Ile Leu Ala Asn Gln Ile Gln Gln His Leu Ile 355 360365 Lys Leu Ile His His Asp Gln Val Gly Phe Ile Pro Gly Met Lys Gly 370375 380 Trp Phe Asn Ile Arg Lys Ser Ile Lys Val Ile His His Ile Asn Arg385 390 395 400 Thr Lys Asp Lys Asn His Met Ile Ile Ser Ile His Ala GluLys Ala 405 410 415 Phe Asp Lys Ile Gln Gln Pro Phe Met Leu Lys Thr ValAsn Lys Leu 420 425 430 Val Ile Asp Gly Thr Tyr Leu Lys Ile Ile Arg AlaIle Tyr Asp Lys 435 440 445 Pro Thr Ala Asn Ile Ile Leu Asn Gly Gln LysLeu Glu Ala Phe Pro 450 455 460 Leu Arg Thr Gly Ile Arg Gln Gly Cys ProLeu Ser Pro Leu Leu Phe 465 470 475 480 Asn Ile Val Leu Glu Val Leu AlaArg Ala Ile Arg Gln Glu Lys Glu 485 490 495 Ile Lys Gly Ile Gln Leu GlyLys Glu Lys Val Lys Leu Ser Leu Phe 500 505 510 Ala Asp Asp Met Ile LeuTyr Leu Glu Asn Pro Ile Val Ser Ala Gln 515 520 525 Asn Leu Leu Lys LeuMet Ser Ser Phe Ser Lys Val Ser Gly Tyr Lys 530 535 540 Ile Asn Val GlnLys Ser Gln Ala Phe Leu Tyr Thr Asn Asn Arg Gln 545 550 555 560 Thr GluSer Gln Met Ser Glu Leu Pro Phe Ala Ile Ala Ser Lys Arg 565 570 575 IleLys Tyr Leu Gly Ile Gln Leu Thr Arg Asp Val Lys Asp Leu Phe 580 585 590Lys Glu Asn Tyr Lys Pro Leu Leu Asn Lys Ile Lys Glu Asp Thr Asn 595 600605 Lys Trp Lys Asn Ile Pro Cys Ser Trp Ile Gly Arg Ile Asn Ile Val 610615 620 Lys Met Ala Ile Met Pro Lys Val Ile Tyr Arg Phe Asn Ala Ile Pro625 630 635 640 Ile Lys Leu Pro Met Thr Phe Phe Thr Glu Leu Glu Lys ThrThr Leu 645 650 655 Lys Phe Ile Trp Asn Gln Lys Arg Ala Arg Ile Ala LysThr Ile Leu 660 665 670 Ser Gln Lys Asn Lys Ala Gly Gly Ile Thr Leu ProAsp Phe Lys Leu 675 680 685 Tyr Tyr Lys Ala Thr Val Thr Lys Thr Ala TrpTyr Trp Tyr Gln Asn 690 695 700 Arg Asp Ile Asp Gln Trp Asn Arg Ile GluPro Leu Glu Leu Ile Pro 705 710 715 720 His Ile Tyr Asn His Leu Ile PheAsp Lys Pro Asp Lys Asn Lys Leu 725 730 735 Trp Gly Lys Asp Ser Leu PheAsn Lys Trp Cys Trp Glu Asn Trp Leu 740 745 750 Ala Ile Cys Arg Lys LeuLys Leu Asn Leu Phe Leu Thr Pro Tyr Thr 755 760 765 Lys Ile Asn Ser ArgTrp Ile Lys Asp Leu Asn Val Arg Pro Lys Thr 770 775 780 Ile Lys Ile LeuGlu Lys Asn Leu Gly Asn Thr Ile Gln Asp Ile Gly 785 790 795 800 Val GlyLys Asp Phe Met Thr Lys Thr Pro Lys Ala Met Ala Thr Lys 805 810 815 AlaLys Ile Asp Lys Trp Asp Ile Ile Lys Leu Lys Ser Phe Cys Thr 820 825 830Ala Lys Glu Thr Thr Ile Ile Val Asn Arg Gln Pro Thr Glu Trp Glu 835 840845 Lys Ile Phe Lys Ile Tyr Pro Ser Asp Lys Gly Leu Ile Ser Arg Ile 850855 860 Tyr Lys Glu Leu Lys Gln Ile Tyr Lys Lys Lys Ser Asn Asn Pro Ile865 870 875 880 Lys Asn Trp Ala Lys Asp Met Asn Arg His Phe Ser Lys GluAsp Ile 885 890 895 Tyr Ala Val Asn Arg His Met Lys Thr Cys Ser Ser LeuLeu Ala Ile 900 905 910 Arg Glu Met Gln Ile Lys Thr Thr Met Arg Tyr HisPhe Thr Pro Val 915 920 925 Arg Met Ala Ser Ile Lys Lys Ser Gly Asn AsnArg 930 935 940 <210> SEQ ID NO 236 <211> LENGTH: 58 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 236 Met Ala Ile Glu Val CysTrp Pro Leu Pro Leu Asp Gly Leu Leu Leu 1 5 10 15 Leu Ala Leu Glu PheLeu Arg Pro Leu Phe Ile Ile Pro Gln Ser Phe 20 25 30 Phe Leu Leu Pro AlaMet Leu Cys Leu Phe Phe Ala Leu Leu Ser Pro 35 40 45 Arg Thr Thr Phe PheHis Phe His Ser Gly 50 55 <210> SEQ ID NO 237 <211> LENGTH: 34 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:UNSURE <222> LOCATION: (28) <400> SEQUENCE: 237 Met Pro Leu His Leu GlyTyr Lys Val Ser Pro Pro Pro Gln Ser His 1 5 10 15 Gly Leu Ala Asn TyrLeu Ser Val Phe Asp Cys Xaa Val Val Ser Thr 20 25 30 Gly Glu <210> SEQID NO 238 <211> LENGTH: 27 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<400> SEQUENCE: 238 Met Arg Lys Val Cys Val Pro Ala Phe Met Thr Ile GluSer Arg Gln 1 5 10 15 Leu Leu Ser Gly Val Ser Ala Cys Phe Gln Gln 20 25<210> SEQ ID NO 239 <211> LENGTH: 26 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: UNSURE <222> LOCATION: (22)<400> SEQUENCE: 239 Met Thr Ser Ile Thr Val Leu Phe Ser Lys Lys Arg LeuSer Leu Met 1 5 10 15 Ala Ser Arg Cys Val Xaa Leu Met Arg Tyr 20 25<210> SEQ ID NO 240 <211> LENGTH: 45 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 240 Met Lys Ser Gln Leu Gln Ser Leu His ProPhe Phe Ser Lys Leu Ala 1 5 10 15 Leu Leu Val Ser Val Leu Phe Tyr IleIle Trp Leu His Leu Thr Val 20 25 30 Phe Lys Lys Ser Ser Val Leu Gln LysAsn Phe Lys Leu 35 40 45 <210> SEQ ID NO 241 <211> LENGTH: 65 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 241 Met Ile GlyIle Thr Trp Cys Phe Glu Leu Ile His Pro Thr Leu Glu 1 5 10 15 Leu ThrAla Thr Val Pro Asp Phe His Arg Tyr Ala Ser Phe His Ser 20 25 30 Gly SerLeu Pro Glu Val Leu His Ser Gly Glu His Ala Gln Val Ser 35 40 45 Pro AlaLeu Gln Asn His Pro Glu Cys Gln Arg Leu Gln His Lys Gly 50 55 60 Lys 65<210> SEQ ID NO 242 <211> LENGTH: 42 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 242 Ile Phe Thr Ala Met Pro Pro Phe Thr LeuGly Val Phe Gln Arg Ser 1 5 10 15 Cys Thr Arg Glu Ser Met Leu Arg PhePro Gln Leu Tyr Lys Ile Thr 20 25 30 Gln Asn Ala Lys Asp Phe Asn Thr ArgVal 35 40 <210> SEQ ID NO 243 <211> LENGTH: 40 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: UNSURE <222>LOCATION: (2) <400> SEQUENCE: 243 Met Xaa Leu Val Leu Leu Thr Arg LeuIle Arg Arg Ser Leu Tyr Thr 1 5 10 15 Lys Arg Asn Leu Leu Ser His SerHis Asn Lys Thr Ser His Gln Thr 20 25 30 Asn Asp Thr Lys Ser Glu Asn His35 40 <210> SEQ ID NO 244 <211> LENGTH: 56 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <400> SEQUENCE: 244 Met Phe Pro Glu Leu Ala SerLeu Tyr Pro Gly Lys Gly Thr Ser Phe 1 5 10 15 Ser Trp Ala Val Pro ProPro Gln Lys Pro Glu Ser Gln Pro Cys Arg 20 25 30 Val Pro Ser Ser Ser PheGln Ile Gln Ile Thr Pro Thr Ser Ser Leu 35 40 45 Gly Ser Pro Ser Leu ArgThr Gln 50 55 <210> SEQ ID NO 245 <211> LENGTH: 26 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <400> SEQUENCE: 245 Met Lys Lys Pro Glu Ala GluAla Ala Leu Thr Leu Arg Asn Pro Val 1 5 10 15 Ser Gln Arg Asp Leu AlaIle Leu Ala Ser 20 25 <210> SEQ ID NO 246 <211> LENGTH: 43 <212> TYPE:PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 246 Met Pro Ile Tyr ProCys Pro Cys Arg Val Gly Arg Lys Asn Leu Met 1 5 10 15 Leu Ala Asn SerPro His Phe Asn Ser Thr Leu Gln Thr Leu Ser Lys 20 25 30 Cys Leu Leu PheVal Arg Gln Tyr Ala Ser His 35 40 <210> SEQ ID NO 247 <211> LENGTH: 49<212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: UNSURE <222> LOCATION: (12)..(33) <400> SEQUENCE: 247 Met LysGln Trp Asp Ala Val Arg Lys Arg Lys Xaa Xaa Xaa Xaa Xaa 1 5 10 15 XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 20 25 30 XaaCys Arg Gly Lys Val Asn Lys Asn Cys Ile Ile Leu Gly Val Phe 35 40 45 Cys<210> SEQ ID NO 248 <211> LENGTH: 24 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 248 Met Pro Tyr Asp Ser Thr Tyr Ile Lys SerLys His Gln Ala Val Leu 1 5 10 15 Ser Met Ile Val Lys Leu Val Gly 20<210> SEQ ID NO 249 <211> LENGTH: 30 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 249 Met His Ile Ser Phe Gly Ile Gln Ile IleVal Asn Asp Gly Glu Leu 1 5 10 15 Thr Ser Asn Ile Ser Ser Tyr Thr ThrAsn Val Ile Lys Pro 20 25 30 <210> SEQ ID NO 250 <211> LENGTH: 192 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:UNSURE <222> LOCATION: (136) <400> SEQUENCE: 250 Met Pro Ser Val Arg AlaHis Pro Asn Pro Arg Ala Glu Gly His Glu 1 5 10 15 Gly Ala Lys Ser LeuArg Asn Ala Ile Leu Arg Leu Val Arg Asp Met 20 25 30 Glu Ile Arg Thr GlnGly Gly Pro Gly Leu Gly Asn Asp Trp Glu Thr 35 40 45 Cys Leu Gly Ser GlnAsp Leu Gly Val Leu Thr Pro Ser Pro His Pro 50 55 60 Ala Val Pro Ser ValPro Ser Pro Ser Leu Ser Lys Pro Leu Gly Ile 65 70 75 80 Glu Trp Pro LeuLeu Phe Trp Cys Pro Gly Val Ile Val Pro Lys Leu 85 90 95 Leu Phe Pro ValPro Ser Pro Gln Arg Leu Val Arg Val Gly Met Arg 100 105 110 Asp Gly GluGly Leu Gly Leu Trp Glu Gln Val Gly Gly Leu Ile Cys 115 120 125 Gly LeuSer Asp Ser Gln Leu Xaa Pro Arg Trp Gly Met Ser Pro Ser 130 135 140 LeuLeu Ser Val Trp Val Arg Lys Thr Gly Cys Asp Pro Glu Glu Gly 145 150 155160 Lys Ile Glu Lys Glu Gly Lys Asp Val Gly Glu Gly Gly Glu Arg Gln 165170 175 Asp Arg Arg Lys Glu Val Glu Glu Glu Val Val Gly Ile Gly Met Arg180 185 190 <210> SEQ ID NO 251 <211> LENGTH: 45 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <400> SEQUENCE: 251 Met Gln Phe Cys Lys Ile LysCys Leu Ser Arg His Ala Tyr Asn Pro 1 5 10 15 Ala Ile Ala Cys Leu GlyAla Tyr Leu Thr Glu Met Asn Ile Tyr Asn 20 25 30 Tyr Ile Ile Ile Cys ThrPro Asn Ser Ser Gln Leu Tyr 35 40 45 <210> SEQ ID NO 252 <211> LENGTH:169 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 252 MetAla Pro Ser Glu Asp Pro Arg Asp Trp Arg Ala Asn Leu Lys Gly 1 5 10 15Thr Ile Arg Glu Thr Gly Leu Glu Thr Ser Ser Gly Gly Lys Leu Ala 20 25 30Gly His Gln Lys Thr Val Pro Thr Ala His Leu Thr Phe Val Ile Asp 35 40 45Cys Thr His Gly Lys Gln Leu Ser Leu Ala Ala Thr Ala Ser Pro Pro 50 55 60Gln Ala Pro Ser Pro Asn Arg Gly Leu Val Thr Pro Pro Met Lys Thr 65 70 7580 Tyr Ile Val Phe Cys Gly Glu Asn Trp Pro His Leu Thr Arg Val Thr 85 9095 Pro Met Gly Gly Gly Cys Leu Ala Gln Ala Arg Ala Thr Leu Pro Leu 100105 110 Cys Arg Gly Ser Val Ala Ser Ala Ser Phe Pro Val Ser Pro Leu Cys115 120 125 Pro Gln Glu Val Pro Glu Ala Lys Gly Lys Pro Val Lys Ala AlaPro 130 135 140 Val Arg Ser Ser Thr Trp Gly Thr Val Lys Asp Ser Leu LysAla Leu 145 150 155 160 Ser Ser Cys Val Cys Gly Gln Ala Asp 165 <210>SEQ ID NO 253 <211> LENGTH: 69 <212> TYPE: PRT <213> ORGANISM: Homosapiens <400> SEQUENCE: 253 Met Phe Asn Val Arg Leu His Gln Asn Met CysGln Leu Thr Met Phe 1 5 10 15 Asn Met Phe His Leu Gln Asn Phe Leu GluGly Lys Lys Ser Phe Leu 20 25 30 Val Asn Met Phe Phe Cys Leu Cys Phe IleIle Leu Ser Thr Met Asp 35 40 45 Thr Gly Asn Gln Ser Thr Val Asn Asn HisArg His His Phe Val Val 50 55 60 Leu Phe Leu Arg Val 65 <210> SEQ ID NO254 <211> LENGTH: 33 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 254 Met Glu Val Arg Ser Val Ile Pro Gln Val Leu Asn Ala TrpAla Ser 1 5 10 15 Leu Met Ser Phe Tyr Gln Leu Ser Ala Thr Cys Val LysPhe His Leu 20 25 30 Ser <210> SEQ ID NO 255 <211> LENGTH: 72 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:UNSURE <222> LOCATION: (59) <221> NAME/KEY: UNSURE <222> LOCATION: (65)<400> SEQUENCE: 255 Met Trp Thr Thr Cys Asn Val Thr Lys Gln Lys Glu ThrGln Glu Ala 1 5 10 15 Asn Ile Pro Ile Tyr Ser Pro Leu Ser Ala Leu ThrGln Gln Asn Lys 20 25 30 Thr Lys Pro Ala Thr Thr Ile Arg Phe Val Lys IleLeu Val Val Arg 35 40 45 Ile Pro Thr Leu Ser Ser Gln Gln Phe Gly Xaa GlnLys Ser Leu Val 50 55 60 Xaa Met Ser Val His Val Lys Ser 65 70 <210> SEQID NO 256 <211> LENGTH: 131 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<220> FEATURE: <221> NAME/KEY: UNSURE <222> LOCATION: (31)..(93) <221>NAME/KEY: UNSURE <222> LOCATION: (111)..(121) <400> SEQUENCE: 256 MetTyr Ala Ser Asn Asn Leu Ser Arg Gly Arg Ile Pro Lys Glu Asn 1 5 10 15Ile Cys Ser Ser Phe Phe Leu Leu Arg Phe Phe Cys Ile Phe Xaa Xaa 20 25 30Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 35 40 45Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 50 55 60Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 65 70 7580 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Thr Val Phe 85 9095 Pro Leu Leu Ser Tyr Asn Asn Gln His Arg Arg Leu Leu Trp Xaa Gln 100105 110 Met Trp Gly Asn Phe Phe His Ala Lys Xaa Ala Val Arg Ala Ala Val115 120 125 Ser Pro Thr 130 <210> SEQ ID NO 257 <211> LENGTH: 44 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 257 Glu Ser PheTyr Asp Thr Phe His Thr Val Ala Asp Met Met Tyr Phe 1 5 10 15 Cys GlnMet Leu Ala Val Val Glu Thr Ile Asn Ala Ala Ile Gly Val 20 25 30 Thr ThrSer Pro Val Leu Pro Ser Leu Ile Gln Val 35 40 <210> SEQ ID NO 258 <211>LENGTH: 70 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: UNSURE <222> LOCATION: (8)..(52) <221> NAME/KEY: UNSURE<222> LOCATION: (57) <400> SEQUENCE: 258 Met Phe Ile Phe Thr Phe His XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 15 Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 20 25 30 Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 35 40 45 Xaa Xaa Xaa Xaa Cys Phe PhePro Xaa Trp Phe Leu Leu Phe Leu Leu 50 55 60 Arg Ser Val Ser Phe Cys 6570 <210> SEQ ID NO 259 <211> LENGTH: 61 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: UNSURE <222> LOCATION:(16)..(53) <400> SEQUENCE: 259 Met Lys Ile Thr Tyr Leu Asp Ile Leu GluLys Tyr Ile His Ser Xaa 1 5 10 15 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa 20 25 30 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa 35 40 45 Xaa Xaa Xaa Xaa Xaa Glu Ser Thr GlnIle Gly Pro Glu 50 55 60 <210> SEQ ID NO 260 <211> LENGTH: 2383 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 260 Met Glu ThrArg Ser Pro Gly Leu Asn Asn Met Lys Pro Gln Ser Leu 1 5 10 15 Gln LeuVal Leu Glu Glu Gln Val Leu Ala Leu Gln Gln Gln Met Ala 20 25 30 Glu AsnGln Ala Ala Ser Trp Arg Lys Leu Lys Asn Ser Gln Glu Ala 35 40 45 Gln GlnArg Gln Ala Thr Leu Val Arg Lys Leu Gln Ala Lys Val Leu 50 55 60 Gln TyrArg Ser Trp Cys Gln Glu Leu Glu Lys Arg Leu Glu Ala Thr 65 70 75 80 GlyGly Pro Ile Pro Gln Arg Trp Glu Asn Val Glu Glu Pro Asn Leu 85 90 95 AspGlu Leu Leu Val Arg Leu Glu Glu Glu Gln Gln Arg Cys Glu Ser 100 105 110Leu Ala Gln Val Asn Thr Gln Leu Arg Leu His Met Glu Lys Ala Asp 115 120125 Val Val Asn Lys Ala Leu Arg Glu Asp Val Glu Lys Leu Thr Val Asp 130135 140 Trp Ser Arg Ala Arg Asp Glu Leu Met Arg Lys Glu Ser Gln Trp Gln145 150 155 160 Met Glu Gln Glu Trp Ser Leu Leu Phe Ser Leu Leu Val LeuArg Asp 165 170 175 Leu Met Glu Leu Lys Ala Glu His Val Arg Leu Ser GlySer Leu Leu 180 185 190 Thr Cys Cys Leu Arg Leu Thr Val Gly Ala Gln SerArg Glu Pro Asn 195 200 205 Gly Ser Gly Arg Met Asn Gly Arg Glu Pro AlaGln Leu Leu Leu Leu 210 215 220 Leu Ala Lys Thr Gln Glu Leu Glu Lys GluAla His Glu Arg Ser Gln 225 230 235 240 Glu Leu Ile Gln Leu Lys Ser GlnGly Asp Leu Glu Lys Ala Glu Leu 245 250 255 Gln Asp Arg Val Thr Glu LeuSer Ala Leu Leu Thr Gln Ser Gln Lys 260 265 270 Gln Asn Glu Asp Tyr GluLys Met Ile Lys Ala Leu Arg Glu Thr Val 275 280 285 Glu Ile Leu Glu ThrAsn His Thr Glu Leu Met Glu His Glu Ala Ser 290 295 300 Leu Ser Arg AsnAla Gln Glu Glu Lys Leu Ser Leu Gln Gln Val Ile 305 310 315 320 Lys AspIle Thr Gln Val Met Val Glu Glu Gly Asp Asn Ile Ala Gln 325 330 335 GlySer Gly His Glu Asn Ser Leu Glu Leu Asp Ser Ser Ile Phe Ser 340 345 350Gln Phe Asp Tyr Gln Asp Ala Asp Lys Ala Leu Thr Leu Val Arg Ser 355 360365 Val Leu Thr Arg Arg Arg Gln Ala Val Gln Asp Leu Arg Gln Gln Leu 370375 380 Ala Gly Cys Gln Glu Ala Val Asn Leu Leu Gln Gln Gln His Asp Gln385 390 395 400 Trp Glu Glu Glu Gly Lys Ala Leu Arg Gln Arg Leu Gln LysLeu Thr 405 410 415 Gly Glu Arg Asp Thr Leu Ala Gly Gln Thr Val Asp LeuGln Gly Glu 420 425 430 Val Asp Ser Leu Ser Lys Glu Arg Glu Leu Leu GlnLys Ala Arg Glu 435 440 445 Glu Leu Arg Gln Gln Leu Glu Val Leu Glu GlnGlu Ala Trp Arg Leu 450 455 460 Arg Arg Val Asn Val Glu Leu Gln Leu GlnGly Asp Ser Ala Gln Gly 465 470 475 480 Gln Lys Glu Glu Gln Gln Glu GluLeu His Leu Ala Val Arg Glu Arg 485 490 495 Glu Arg Leu Gln Glu Met LeuMet Gly Leu Glu Ala Lys Gln Ser Glu 500 505 510 Ser Leu Ser Glu Leu IleThr Leu Arg Glu Ala Leu Glu Ser Ser His 515 520 525 Leu Glu Gly Glu LeuLeu Arg Gln Glu Gln Thr Glu Val Thr Ala Ala 530 535 540 Leu Ala Arg AlaGlu Gln Ser Ile Ala Glu Leu Ser Ser Ser Glu Asn 545 550 555 560 Thr LeuLys Thr Glu Val Ala Asp Leu Arg Ala Ala Ala Val Lys Leu 565 570 575 SerAla Leu Asn Glu Ala Leu Ala Leu Asp Lys Val Gly Leu Asn Gln 580 585 590Gln Leu Leu Gln Leu Glu Glu Glu Asn Gln Ser Val Cys Ser Arg Met 595 600605 Glu Ala Ala Glu Gln Ala Arg Asn Ala Leu Gln Val Asp Leu Ala Glu 610615 620 Ala Glu Lys Arg Arg Glu Ala Leu Trp Glu Lys Asn Thr His Leu Glu625 630 635 640 Ala Gln Leu Gln Lys Ala Glu Glu Ala Gly Ala Glu Leu GlnAla Asp 645 650 655 Leu Arg Asp Ile Gln Glu Glu Lys Glu Glu Ile Gln LysLys Leu Ser 660 665 670 Glu Ser Arg His Gln Gln Glu Ala Ala Thr Thr GlnLeu Glu Gln Leu 675 680 685 His Gln Glu Ala Lys Arg Gln Glu Glu Val LeuAla Arg Ala Val Gln 690 695 700 Glu Lys Glu Ala Leu Val Arg Glu Lys AlaAla Leu Glu Val Arg Leu 705 710 715 720 Gln Ala Val Glu Arg Asp Arg GlnAsp Leu Ala Glu Gln Leu Gln Gly 725 730 735 Leu Ser Ser Ala Lys Glu LeuLeu Glu Ser Ser Leu Phe Glu Ala Gln 740 745 750 Gln Gln Asn Ser Val IleGlu Val Thr Lys Gly Gln Leu Glu Val Gln 755 760 765 Ile Gln Thr Val ThrGln Ala Lys Glu Val Ile Gln Gly Glu Val Arg 770 775 780 Cys Leu Lys LeuGlu Leu Asp Thr Glu Arg Ser Gln Ala Glu Gln Glu 785 790 795 800 Arg AspAla Ala Ala Arg Gln Leu Ala Gln Ala Glu Gln Glu Gly Lys 805 810 815 ThrAla Leu Glu Gln Gln Lys Ala Ala His Glu Lys Glu Val Asn Gln 820 825 830Leu Arg Glu Lys Trp Glu Lys Glu Arg Ser Trp His Gln Gln Glu Leu 835 840845 Ala Lys Ala Leu Glu Ser Leu Glu Arg Glu Lys Met Glu Leu Glu Met 850855 860 Arg Leu Lys Glu Gln Gln Thr Glu Met Glu Ala Ile Gln Ala Gln Arg865 870 875 880 Glu Glu Glu Arg Thr Gln Ala Glu Ser Ala Leu Cys Gln MetGln Leu 885 890 895 Glu Thr Glu Lys Glu Arg Val Ser Leu Leu Glu Thr LeuLeu Gln Thr 900 905 910 Gln Lys Glu Leu Ala Asp Ala Ser Gln Gln Leu GluArg Leu Arg Gln 915 920 925 Asp Met Lys Val Gln Lys Leu Lys Glu Gln GluThr Thr Gly Ile Leu 930 935 940 Gln Thr Gln Leu Gln Glu Ala Gln Arg GluLeu Lys Glu Ala Ala Arg 945 950 955 960 Gln His Arg Asp Asp Leu Ala AlaLeu Gln Glu Glu Ser Ser Ser Leu 965 970 975 Leu Gln Asp Lys Met Asp LeuGln Lys Gln Val Glu Asp Leu Lys Ser 980 985 990 Gln Leu Val Ala Gln AspAsp Ser Gln Arg Leu Val Glu Gln Glu Val 995 1000 1005 Gln Glu Lys LeuArg Glu Thr Gln Glu Tyr Asn Arg Ile Gln Lys Glu 1010 1015 1020 Leu GluArg Glu Lys Ala Ser Leu Thr Leu Ser Leu Met Glu Lys Glu 1025 1030 10351040 Gln Arg Leu Leu Val Leu Gln Glu Ala Asp Ser Ile Arg Gln Gln Glu1045 1050 1055 Leu Ser Ala Leu Arg Gln Asp Met Gln Glu Ala Gln Gly GluGln Lys 1060 1065 1070 Glu Leu Ser Ala Gln Met Glu Leu Leu Arg Gln GluVal Lys Glu Lys 1075 1080 1085 Glu Ala Asp Phe Leu Ala Gln Glu Ala GlnLeu Leu Glu Glu Leu Glu 1090 1095 1100 Ala Ser His Ile Thr Glu Gln GlnLeu Arg Ala Ser Leu Trp Ala Gln 1105 1110 1115 1120 Glu Ala Lys Ala AlaGln Leu Gln Leu Arg Leu Arg Ser Thr Glu Ser 1125 1130 1135 Gln Leu GluAla Leu Ala Ala Glu Gln Gln Pro Gly Asn Gln Ala Gln 1140 1145 1150 AlaGln Ala Gln Leu Ala Ser Leu Tyr Ser Ala Leu Gln Gln Ala Leu 1155 11601165 Gly Ser Val Cys Glu Ser Arg Pro Glu Leu Ser Gly Gly Gly Asp Ser1170 1175 1180 Ala Pro Ser Val Trp Gly Leu Glu Pro Asp Gln Asn Gly AlaArg Ser 1185 1190 1195 1200 Leu Phe Lys Arg Gly Pro Leu Leu Thr Ala LeuSer Ala Glu Ala Val 1205 1210 1215 Ala Ser Ala Leu His Lys Leu His GlnAsp Leu Trp Lys Thr Gln Gln 1220 1225 1230 Thr Arg Asp Val Leu Arg AspGln Val Gln Lys Leu Glu Glu Arg Leu 1235 1240 1245 Thr Asp Thr Glu AlaGlu Lys Ser Gln Val His Thr Glu Leu Gln Asp 1250 1255 1260 Leu Gln ArgGln Leu Ser Gln Asn Gln Glu Glu Lys Ser Lys Trp Glu 1265 1270 1275 1280Gly Lys Gln Asn Ser Leu Glu Ser Glu Leu Met Glu Leu His Glu Thr 12851290 1295 Met Ala Ser Leu Gln Ser Arg Leu Arg Arg Ala Glu Leu Gln ArgMet 1300 1305 1310 Glu Ala Gln Gly Glu Arg Glu Leu Leu Gln Ala Ala LysGlu Asn Leu 1315 1320 1325 Thr Ala Gln Val Glu His Leu Gln Ala Ala ValVal Glu Ala Arg Ala 1330 1335 1340 Gln Ala Ser Ala Ala Gly Ile Leu GluGlu Asp Leu Arg Thr Ala Arg 1345 1350 1355 1360 Ser Ala Leu Lys Leu LysAsn Glu Glu Val Glu Ser Glu Arg Glu Arg 1365 1370 1375 Ala Gln Ala LeuGln Glu Gln Gly Glu Leu Lys Val Ala Gln Gly Lys 1380 1385 1390 Ala LeuGln Glu Asn Leu Ala Leu Leu Thr Gln Thr Leu Ala Glu Arg 1395 1400 1405Glu Glu Glu Val Glu Thr Leu Arg Gly Gln Ile Gln Glu Leu Glu Lys 14101415 1420 Gln Arg Glu Met Gln Lys Ala Ala Leu Glu Leu Leu Ser Leu AspLeu 1425 1430 1435 1440 Lys Lys Arg Asn Gln Glu Val Asp Leu Gln Gln GluGln Ile Gln Glu 1445 1450 1455 Leu Glu Lys Cys Arg Ser Val Leu Glu HisLeu Pro Met Ala Val Gln 1460 1465 1470 Glu Arg Glu Gln Lys Leu Thr ValGln Arg Glu Gln Ile Arg Glu Leu 1475 1480 1485 Glu Lys Asp Arg Glu ThrGln Arg Asn Val Leu Glu His Gln Leu Leu 1490 1495 1500 Glu Leu Glu LysLys Asp Gln Met Ile Glu Ser Gln Arg Gly Gln Val 1505 1510 1515 1520 GlnAsp Leu Lys Lys Gln Leu Val Thr Leu Glu Cys Leu Ala Leu Glu 1525 15301535 Leu Glu Glu Asn His His Lys Met Glu Cys Gln Gln Lys Leu Ile Lys1540 1545 1550 Glu Leu Glu Gly Gln Arg Glu Thr Gln Arg Val Ala Leu ThrHis Leu 1555 1560 1565 Thr Leu Asp Leu Glu Glu Arg Ser Gln Glu Leu GlnAla Gln Ser Ser 1570 1575 1580 Gln Ile His Asp Leu Glu Ser His Ser ThrVal Leu Ala Arg Glu Leu 1585 1590 1595 1600 Gln Glu Arg Asp Gln Glu ValLys Ser Gln Arg Glu Gln Ile Glu Glu 1605 1610 1615 Leu Gln Arg Gln LysGlu His Leu Thr Gln Asp Leu Glu Arg Arg Asp 1620 1625 1630 Gln Glu LeuMet Leu Gln Lys Glu Arg Ile Gln Val Leu Glu Asp Gln 1635 1640 1645 ArgThr Arg Gln Thr Lys Ile Leu Glu Glu Asp Leu Glu Gln Ile Lys 1650 16551660 Leu Ser Leu Arg Glu Arg Gly Arg Glu Leu Thr Thr Gln Arg Gln Leu1665 1670 1675 1680 Met Gln Glu Arg Ala Glu Glu Gly Lys Gly Pro Ser LysAla Gln Arg 1685 1690 1695 Gly Ser Leu Glu His Met Lys Leu Ile Leu ArgAsp Lys Glu Lys Glu 1700 1705 1710 Val Glu Cys Gln Gln Glu His Ile HisGlu Leu Gln Glu Leu Lys Asp 1715 1720 1725 Gln Leu Glu Gln Gln Leu GlnGly Leu His Arg Lys Val Gly Glu Thr 1730 1735 1740 Ser Leu Leu Leu SerGln Arg Glu Gln Glu Ile Val Val Leu Gln Gln 1745 1750 1755 1760 Gln LeuGln Glu Ala Arg Glu Gln Gly Glu Leu Lys Glu Gln Ser Leu 1765 1770 1775Gln Ser Gln Leu Asp Glu Ala Gln Arg Ala Leu Ala Gln Arg Asp Gln 17801785 1790 Glu Leu Glu Ala Leu Gln Gln Glu Gln Gln Gln Ala Gln Gly GlnGlu 1795 1800 1805 Glu Arg Val Lys Glu Lys Ala Asp Ala Leu Gln Gly AlaLeu Glu Gln 1810 1815 1820 Ala His Met Thr Leu Lys Glu Arg His Gly GluLeu Gln Asp His Lys 1825 1830 1835 1840 Glu Gln Ala Arg Arg Leu Glu GluGlu Leu Ala Val Glu Gly Arg Arg 1845 1850 1855 Val Gln Ala Leu Glu GluVal Leu Gly Asp Leu Arg Ala Glu Ser Arg 1860 1865 1870 Glu Gln Glu LysAla Leu Leu Ala Leu Gln Gln Gln Cys Ala Glu Gln 1875 1880 1885 Ala GlnGlu His Glu Val Glu Thr Arg Ala Leu Gln Asp Ser Trp Leu 1890 1895 1900Gln Ala Gln Ala Val Leu Lys Glu Arg Asp Gln Glu Leu Glu Ala Leu 19051910 1915 1920 Arg Ala Glu Ser Gln Ser Ser Arg His Gln Glu Glu Ala AlaArg Ala 1925 1930 1935 Arg Ala Glu Ala Leu Gln Glu Ala Leu Gly Lys AlaHis Ala Ala Leu 1940 1945 1950 Gln Gly Lys Glu Gln His Leu Leu Glu GlnAla Glu Leu Ser Arg Ser 1955 1960 1965 Leu Glu Ala Ser Thr Ala Thr LeuGln Ala Ser Leu Asp Ala Cys Gln 1970 1975 1980 Ala His Ser Arg Gln LeuGlu Glu Ala Leu Arg Ile Gln Glu Gly Glu 1985 1990 1995 2000 Ile Gln AspGln Asp Leu Arg Tyr Gln Glu Asp Val Gln Gln Leu Gln 2005 2010 2015 GlnAla Leu Ala Gln Arg Asp Glu Glu Leu Arg His Gln Gln Glu Arg 2020 20252030 Glu Gln Leu Leu Glu Lys Ser Leu Ala Gln Arg Val Gln Glu Asn Met2035 2040 2045 Ile Gln Glu Lys Gln Asn Leu Gly Gln Glu Arg Glu Glu GluGlu Ile 2050 2055 2060 Arg Gly Leu His Gln Ser Val Arg Glu Leu Gln LeuThr Leu Ala Gln 2065 2070 2075 2080 Lys Glu Gln Glu Ile Leu Glu Leu ArgGlu Thr Gln Gln Arg Asn Asn 2085 2090 2095 Leu Glu Ala Leu Pro His SerHis Lys Thr Ser Pro Met Glu Glu Gln 2100 2105 2110 Ser Leu Lys Leu AspSer Leu Glu Pro Arg Leu Gln Arg Glu Leu Glu 2115 2120 2125 Arg Leu GlnAla Ala Leu Arg Gln Thr Glu Ala Arg Glu Ile Glu Trp 2130 2135 2140 ArgGlu Lys Ala Gln Asp Leu Ala Leu Ser Leu Ala Gln Thr Lys Ala 2145 21502155 2160 Ser Val Ser Ser Leu Gln Glu Val Ala Met Phe Leu Gln Ala SerVal 2165 2170 2175 Leu Glu Arg Asp Ser Glu Gln Gln Arg Leu Gln Asp GluLeu Glu Leu 2180 2185 2190 Thr Arg Arg Ala Leu Glu Lys Glu Arg Leu HisSer Pro Gly Ala Thr 2195 2200 2205 Ser Thr Ala Glu Leu Gly Ser Arg GlyGlu Gln Gly Val Gln Leu Gly 2210 2215 2220 Glu Val Ser Gly Val Glu AlaGlu Pro Ser Pro Asp Gly Met Glu Lys 2225 2230 2235 2240 Gln Ser Trp ArgGln Arg Leu Glu His Leu Gln Gln Ala Val Ala Arg 2245 2250 2255 Leu GluIle Asp Arg Ser Arg Leu Gln Arg His Asn Val Gln Leu Arg 2260 2265 2270Ser Thr Leu Glu Gln Asp Gly Arg Gly Gln Lys Asn Ser Asp Ala Lys 22752280 2285 Cys Val Ala Glu Leu Gln Lys Glu Val Val Leu Leu Gln Ala GlnLeu 2290 2295 2300 Thr Leu Glu Arg Lys Gln Lys Gln Asp Tyr Ile Thr ArgSer Ala Gln 2305 2310 2315 2320 Thr Ser Arg Glu Leu Ala Gly Leu His HisSer Leu Ser His Ser Leu 2325 2330 2335 Leu Ala Val Ala Gln Ala Pro GluAla Thr Val Leu Glu Ala Glu Thr 2340 2345 2350 Arg Arg Leu Asp Glu SerLeu Thr Gln Ser Leu Thr Ser Pro Gly Pro 2355 2360 2365 Val Leu Leu HisPro Ser Pro Ser Thr Thr Gln Ala Ala Ser Arg 2370 2375 2380 <210> SEQ IDNO 261 <211> LENGTH: 43 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<400> SEQUENCE: 261 Met Tyr Arg Leu Ile Leu Phe Arg Asn Asn Ser Val LeuGlu Phe Ile 1 5 10 15 Lys Asn Ser Val Ile Ala Phe Ile Pro Lys Cys LeuThr Leu Pro Thr 20 25 30 Ala Ser His Lys Ser Ile Tyr Phe Lys Ala Phe 3540 <210> SEQ ID NO 262 <211> LENGTH: 34 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 262 Met Asp Pro Asn Phe Asp Ile Val His ThrVal Phe Ile Leu Cys Met 1 5 10 15 Glu Leu Ile Thr Asp Phe Ala Cys LysGlu Arg Ile Val Cys Leu Asn 20 25 30 Phe Val <210> SEQ ID NO 263 <211>LENGTH: 78 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:263 Met Met Glu Asn Ser Ala Pro Asn Ser Leu Met Asn Lys Glu Met Asp 1 510 15 His Leu Met Asp Glu Gly Val Gln Arg Thr Arg Val Ala Leu Gly Gln 2025 30 Trp Leu Val Ala Ala Val Ile Gln Asp Leu Gly Ser Val Leu Cys Pro 3540 45 Leu Pro Pro Ser Val Leu Ala Ser Arg Trp Gln Gly Val Ser Phe Pro 5055 60 Glu Ser His Gln Leu Arg Gln Asn Pro Glu Ala Gly Lys Thr 65 70 75<210> SEQ ID NO 264 <211> LENGTH: 85 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: UNSURE <222> LOCATION:(15)..(72) <400> SEQUENCE: 264 Met Gly Ile Tyr Ile Ile Tyr Ser Pro ArgThr Val Ile Arg Xaa Xaa 1 5 10 15 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa 20 25 30 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa 35 40 45 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa 50 55 60 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa LeuLeu Ala Ser Phe Cys Phe Pro 65 70 75 80 Leu Val Leu Gly Phe 85 <210> SEQID NO 265 <211> LENGTH: 471 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<400> SEQUENCE: 265 Leu Ser Phe Gln Ser Gly Asn Ile Ile Val Ala Thr ProGly Arg Leu 1 5 10 15 Glu Asp Met Phe Arg Arg Lys Ala Glu Gly Leu AspLeu Ala Ser Cys 20 25 30 Val Arg Ser Leu Asp Val Leu Val Leu Asp Glu AlaAsp Arg Leu Leu 35 40 45 Asp Met Gly Phe Glu Ala Ser Ile Asn Thr Ile LeuGlu Phe Leu Pro 50 55 60 Lys Gln Arg Arg Thr Gly Leu Phe Ser Ala Thr GlnThr Gln Glu Val 65 70 75 80 Glu Asn Leu Val Arg Ala Gly Leu Arg Asn ProVal Arg Val Ser Val 85 90 95 Lys Glu Lys Gly Val Ala Ala Ser Ser Ala GlnLys Thr Pro Ser Arg 100 105 110 Leu Glu Asn Tyr Tyr Met Val Cys Lys AlaAsp Glu Lys Phe Asn Gln 115 120 125 Leu Val His Phe Leu Arg Asn His LysGln Glu Lys His Leu Val Phe 130 135 140 Phe Gly Thr Cys Ala Cys Val GluTyr Tyr Gly Lys Ala Leu Glu Val 145 150 155 160 Leu Val Lys Gly Val LysIle Met Cys Ile His Gly Lys Met Lys Tyr 165 170 175 Lys Arg Asn Lys IlePhe Met Glu Phe Arg Lys Leu Gln Gly Gly Ile 180 185 190 Leu Val Cys ThrAsp Val Met Ala Arg Gly Ile Asp Ile Pro Glu Val 195 200 205 Asn Trp ValLeu Gln Tyr Asp Pro Pro Ser Asn Ala Ser Ala Phe Val 210 215 220 His ArgCys Gly Arg Thr Ala Arg Ile Gly His Gly Gly Ser Ala Leu 225 230 235 240Val Phe Leu Leu Pro Met Glu Glu Ser Tyr Ile Asn Phe Leu Ala Ile 245 250255 Asn Gln Lys Cys Pro Leu Gln Glu Met Lys Pro Gln Arg Asn Thr Ala 260265 270 Asp Leu Leu Pro Lys Leu Lys Ser Met Ala Leu Ala Asp Arg Ala Val275 280 285 Phe Glu Lys Gly Met Lys Ala Phe Val Ser Tyr Val Gln Ala TyrAla 290 295 300 Lys His Glu Cys Asn Leu Ile Phe Arg Leu Lys Asp Leu AspPhe Ala 305 310 315 320 Ser Leu Ala Arg Gly Phe Ala Leu Leu Arg Met ProLys Met Pro Glu 325 330 335 Leu Arg Gly Lys Gln Phe Pro Asp Phe Val ProVal Asp Val Asn Thr 340 345 350 Asp Thr Ile Pro Phe Lys Asp Lys Ile ArgGlu Lys Gln Arg Gln Lys 355 360 365 Leu Leu Glu Gln Gln Arg Arg Glu LysThr Glu Asn Glu Gly Arg Arg 370 375 380 Lys Phe Ile Lys Asn Lys Ala TrpSer Lys Gln Lys Ala Lys Lys Glu 385 390 395 400 Lys Lys Lys Lys Met AsnGlu Lys Arg Lys Arg Glu Glu Gly Ser Asp 405 410 415 Ile Glu Asp Glu AspMet Glu Glu Leu Leu Asn Asp Thr Arg Leu Leu 420 425 430 Lys Lys Leu LysLys Gly Lys Ile Thr Glu Glu Glu Phe Glu Lys Gly 435 440 445 Leu Leu ThrThr Gly Lys Arg Thr Ile Lys Thr Val Asp Leu Gly Ile 450 455 460 Ser AspLeu Glu Asp Asp Cys 465 470 <210> SEQ ID NO 266 <211> LENGTH: 20 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 266 Met Met ThrSer Leu Ser Tyr Ser Ser Gln Ser Trp Lys Pro Cys Ser 1 5 10 15 Gln SerPhe Lys 20 <210> SEQ ID NO 267 <211> LENGTH: 27 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <400> SEQUENCE: 267 Met Val Phe Leu Glu Ile IlePhe Cys Pro Met Tyr Ser Ile Phe Ile 1 5 10 15 His Thr Gly Phe Ile MetIle Ile Ile Ser Lys 20 25 <210> SEQ ID NO 268 <211> LENGTH: 55 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 268 Met Leu ArgGly Asp Leu Pro Gly Ser Val Leu Pro Leu Ser Leu Arg 1 5 10 15 Leu AsnGly Ala Pro Pro Arg Leu Leu Pro Gly Lys Lys His Ser Gly 20 25 30 Gln AlaGly Pro Glu Pro Val Ser Val Arg Gly Pro Val Ala Cys Pro 35 40 45 Gly GlyArg Ser Leu Gln Gly 50 55 <210> SEQ ID NO 269 <211> LENGTH: 38 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 269 Met Asn GluAla Asn Arg Leu Phe Phe Val Ser Leu Thr Pro Arg Asn 1 5 10 15 Ile MetIle Pro Tyr Lys Ile Leu Ile His Thr His Asp Gln Tyr Phe 20 25 30 Ile ProThr Glu Thr Val 35 <210> SEQ ID NO 270 <211> LENGTH: 71 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 270 Met Leu Thr Leu Val TyrLeu Val Val Glu Asn Gly Leu Leu Pro Leu 1 5 10 15 Phe Pro Glu Leu ThrLeu Phe Pro Leu Ala Arg Arg Ser Gly Gln Arg 20 25 30 Glu Pro Arg Thr GluVal Pro Thr Thr Gln Gln Ala Leu Ser Ser Pro 35 40 45 Leu Thr Ser Asn ValCys Ile His Phe Gln Pro Leu Thr Asp Leu Val 50 55 60 Phe Gln Cys Ile IleIle Leu 65 70 <210> SEQ ID NO 271 <211> LENGTH: 65 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <400> SEQUENCE: 271 Met Glu Glu Ser Lys Ala GlnArg Arg Arg Glu Thr Thr Trp Ser Val 1 5 10 15 Ser Leu Ser Gln Leu IleGln His Pro Thr Asn His Pro Ser His Ser 20 25 30 Leu Ser Ile Ser Leu ValAsn Trp Ser Thr Ile Cys Asn Cys Ser Gln 35 40 45 Val Pro Pro Asn Ser LeuCys Arg Tyr Phe Ser Cys Val Phe His Ser 50 55 60 Leu 65 <210> SEQ ID NO272 <211> LENGTH: 25 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 272 Met Val Pro Ile Ile Ser Tyr Val Lys Met Ser Cys Tyr GluLys Leu 1 5 10 15 Phe Leu Phe Gln Ser Cys Gln Cys Gln 20 25 <210> SEQ IDNO 273 <211> LENGTH: 13 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<400> SEQUENCE: 273 Met Leu Leu Ser Tyr Ser Ala Gln Glu Tyr Leu Ser Lys1 5 10 <210> SEQ ID NO 274 <211> LENGTH: 73 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <400> SEQUENCE: 274 Met Lys Cys Val Ser Glu HisGln Arg Pro Ser Ile Leu Pro Leu Pro 1 5 10 15 Phe Leu Val Val Tyr LysAsn Ser Arg Leu Glu Glu Phe Arg Phe Val 20 25 30 Ala His Phe Phe Pro GlnHis Phe Phe Leu Leu Phe Phe Lys Met Tyr 35 40 45 Cys Leu Phe Pro His SerVal Thr Leu Asp Ile Gly Ile Phe Asn Cys 50 55 60 Val Ile Phe Cys Cys LysLys Gly Lys 65 70 <210> SEQ ID NO 275 <211> LENGTH: 465 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 275 Met Leu Gly Ser Met AlaArg Lys Lys Pro Arg Asn Thr Ser Arg Leu 1 5 10 15 Pro Leu Ala Leu AsnPro Leu Lys Ser Lys Asp Val Leu Ala Val Leu 20 25 30 Ala Glu Arg Asn GluAla Ile Val Pro Val Gly Ala Trp Val Glu Pro 35 40 45 Ala Ser Pro Gly SerSer Glu Ile Pro Ala Tyr Thr Ser Ala Tyr Leu 50 55 60 Ile Glu Glu Glu LeuLys Glu Gln Leu Arg Lys Lys Gln Glu Ala Leu 65 70 75 80 Lys His Phe GlnLys Gln Val Lys Tyr Arg Val Asn Gln Gln Ile Arg 85 90 95 Leu Arg Lys LysGln Gln Leu Gln Lys Ser Tyr Glu Arg Ala Gln Lys 100 105 110 Glu Gly SerIle Ala Met Gln Ser Ser Ala Thr His Leu Thr Ser Lys 115 120 125 Arg ThrSer Val Phe Pro Asn Asn Leu Asn Val Ala Ile Gly Ser Ser 130 135 140 ArgLeu Pro Pro Ser Leu Met Pro Gly Asp Gly Ile Glu Asp Glu Glu 145 150 155160 Asn Gln Asn Glu Leu Phe Gln Gln Gln Ala Gln Ala Leu Ser Glu Thr 165170 175 Met Lys Gln Ala Arg His Arg Leu Ala Ser Phe Lys Thr Val Ile Lys180 185 190 Lys Lys Gly Ser Val Phe Pro Asp Asp Gly Arg Lys Ser Phe LeuThr 195 200 205 Arg Glu Glu Val Leu Ser Arg Lys Pro Ala Ser Thr Gly IleAsn Thr 210 215 220 Gly Ile Arg Gly Glu Leu Pro Ile Lys Val His Gln GlyLeu Leu Ala 225 230 235 240 Ala Val Pro Tyr Gln Asn Tyr Met Glu Asn GlnGlu Leu Asp Tyr Glu 245 250 255 Glu Pro Asp Tyr Glu Glu Ser Ser Ser LeuVal Thr Asp Glu Lys Gly 260 265 270 Lys Glu Asp Leu Phe Gly Arg Gly GlnGln Asp Gln Gln Ala Ile His 275 280 285 Ser Glu Asp Lys Asn Lys Pro PheSer Arg Val Gln Lys Val Lys Phe 290 295 300 Lys Asn Pro Leu Phe Val LeuMet Glu Glu Glu Glu Gln Lys Gln Leu 305 310 315 320 His Phe Glu Gly LeuGln Asp Ile Leu Pro Glu Ala Gln Asp Tyr Phe 325 330 335 Leu Glu Ala GlnGly Asp Leu Leu Glu Thr Gln Gly Asp Leu Thr Gly 340 345 350 Ile Gln SerVal Lys Pro Asp Thr Gln Ala Val Glu Met Lys Val Gln 355 360 365 Val ThrGlu Pro Glu Gly Gln Ala Ile Glu Pro Glu Gly Gln Pro Ile 370 375 380 LysThr Glu Thr Gln Gly Ile Met Leu Lys Ala Gln Ser Ile Glu Leu 385 390 395400 Glu Glu Gly Ser Ile Val Leu Lys Thr Gln Asp Phe Leu Pro Thr Asn 405410 415 Gln Ala Leu Leu Thr Lys Asn Gln Asp Val Leu Leu Lys Asp His Cys420 425 430 Val Leu Pro Lys Asp Gln Ser Ile Leu Leu Lys Tyr Gln Asp GlnAsp 435 440 445 Phe Leu Pro Arg Asp Gln His Val Leu His Lys Asp Gln AspIle Leu 450 455 460 Pro 465 <210> SEQ ID NO 276 <211> LENGTH: 38 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 276 Met Asn LysGln Lys Ile Lys Met Phe Arg Met Lys Ile Leu Leu Lys 1 5 10 15 Trp SerLeu Glu Ile Thr Val Met Ser Ala Leu Gly Ile Glu Ser Arg 20 25 30 Ile AsnSer Gln Ile Pro 35 <210> SEQ ID NO 277 <211> LENGTH: 170 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 277 Met Asp Ile Glu Arg GluGln Val Lys Glu Gln Gln Arg Gln Lys Glu 1 5 10 15 Gln Lys Lys Lys IleGlu Lys Ile Lys Lys Lys Arg Glu Gln Glu Cys 20 25 30 Tyr Ala Ala Glu GlnArg Ile Leu Arg Met Asn Phe His Glu Asp Pro 35 40 45 Tyr Ser Gly Glu LysLeu Ser Glu Ile Leu Ala Gln Leu Gln Leu Gln 50 55 60 Glu Ile Lys Gly ThrArg Glu Lys Gln Gln Arg Glu Lys Glu Tyr Leu 65 70 75 80 Arg Tyr Val GluAla Leu Arg Ala Gln Ile Gln Glu Lys Met Gln Leu 85 90 95 Tyr Asn Ile ThrLeu Pro Pro Leu Cys Cys Cys Gly Pro Asp Phe Trp 100 105 110 Asp Ala HisPro Asp Thr Cys Ala Asn Asn Cys Ile Phe Tyr Lys Asn 115 120 125 His ArgAla Tyr Thr Arg Ala Leu His Ser Phe Ile Asn Ser Cys Asp 130 135 140 ValPro Gly Gly Asn Ser Thr Leu Arg Val Ala Ile His Asn Phe Ala 145 150 155160 Ser Ala His Arg Arg Thr Leu Lys Asn Leu 165 170 <210> SEQ ID NO 278<211> LENGTH: 173 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 278 Ala Tyr Asp Arg Tyr Gln Ser Gly Leu Ser Thr Glu Phe GlnAla Pro 1 5 10 15 Leu Ala Phe Gln Ser Asp Val Asp Lys Glu Glu Asp LysLys Glu Arg 20 25 30 Gln Lys Gln Tyr Leu Arg His Arg Arg Leu Phe Met AspIle Glu Arg 35 40 45 Glu Gln Val Lys Glu Gln Gln Arg Gln Lys Glu Gln LysLys Lys Ile 50 55 60 Glu Lys Ile Lys Lys Lys Arg Glu Gln Glu Cys Tyr AlaAla Glu Gln 65 70 75 80 Arg Ile Leu Arg Met Asn Phe His Glu Asp Pro TyrSer Gly Glu Lys 85 90 95 Leu Ser Glu Ile Leu Ala Gln Leu Gln Leu Gln GluIle Lys Gly Thr 100 105 110 Arg Glu Lys Gln Gln Arg Glu Lys Glu Tyr LeuArg Tyr Val Glu Ala 115 120 125 Leu Arg Ala Gln Ile Gln Glu Lys Met GlnLeu Tyr Asn Ile Thr Leu 130 135 140 Pro Pro Leu Cys Cys Cys Gly Pro AspPhe Trp Asp Ala His Pro Asp 145 150 155 160 Thr Cys Ala Asn Asn Cys IlePhe Tyr Lys Asn His Arg 165 170 <210> SEQ ID NO 279 <211> LENGTH: 15<212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 279 Met IleSer Arg Ile Leu Pro Phe Ile Tyr Ser Thr Ser Ile Arg 1 5 10 15 <210> SEQID NO 280 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<220> FEATURE: <221> NAME/KEY: UNSURE <222> LOCATION: (8) <400>SEQUENCE: 280 Met Asp Thr Gly Leu Phe Phe Xaa Gly Ala Gly 1 5 10 <210>SEQ ID NO 281 <211> LENGTH: 86 <212> TYPE: PRT <213> ORGANISM: Homosapiens <400> SEQUENCE: 281 Met Ala Val Ser Leu Phe Leu Ser Ala Asp ProSer Met Thr Leu Ile 1 5 10 15 Arg Phe Pro Phe Ser Tyr Asn Ser Cys ProTrp Ile Gln Trp Pro Ser 20 25 30 Phe Phe Ser Phe Ala Leu Phe Ser Val ThrVal His His Ile Phe Tyr 35 40 45 Thr Ala Val Asp Val Ile Tyr Ser Asn AspVal Pro Val Pro Phe Val 50 55 60 Cys Leu Phe Leu Glu Thr Pro Ser Gly AlaPhe His Leu Pro Gly Ser 65 70 75 80 Asn Leu Asp Trp Leu Leu 85 <210> SEQID NO 282 <211> LENGTH: 1339 <212> TYPE: PRT <213> ORGANISM: Homosapiens <400> SEQUENCE: 282 Met Ala Val Tyr Cys Tyr Ala Leu Asn Ser LeuVal Ile Met Asn Ser 1 5 10 15 Ala Asn Glu Met Lys Ser Gly Gly Gly ProGly Pro Ser Gly Ser Glu 20 25 30 Thr Pro Pro Pro Pro Arg Arg Ala Val LeuSer Pro Gly Ser Val Phe 35 40 45 Ser Pro Gly Arg Gly Ala Ser Phe Leu PhePro Pro Ala Glu Ser Leu 50 55 60 Ser Pro Glu Glu Pro Arg Ser Pro Gly GlyTrp Arg Ser Gly Arg Arg 65 70 75 80 Arg Leu Asn Ser Ser Ser Gly Ser GlySer Gly Ser Ser Gly Ser Ser 85 90 95 Val Ser Ser Pro Ser Trp Ala Gly ArgLeu Arg Gly Asp Arg Gln Gln 100 105 110 Val Val Ala Ala Gly Thr Leu SerPro Pro Gly Pro Glu Glu Ala Lys 115 120 125 Arg Lys Leu Arg Ile Leu GlnArg Glu Leu Gln Asn Val Gln Val Asn 130 135 140 Gln Lys Val Gly Met PheGlu Ala His Ile Gln Ala Gln Ser Ser Ala 145 150 155 160 Ile Gln Ala ProArg Ser Pro Arg Leu Gly Arg Ala Arg Ser Pro Ser 165 170 175 Pro Cys ProPhe Arg Ser Ser Ser Gln Pro Pro Gly Arg Val Leu Val 180 185 190 Gln GlyAla Arg Ser Glu Glu Arg Arg Thr Lys Ser Trp Gly Glu Gln 195 200 205 CysPro Glu Thr Ser Gly Thr Asp Ser Gly Arg Lys Gly Gly Pro Ser 210 215 220Leu Cys Ser Ser Gln Val Lys Lys Gly Met Pro Pro Leu Pro Gly Arg 225 230235 240 Ala Ala Pro Thr Gly Ser Glu Ala Gln Gly Pro Ser Ala Phe Val Arg245 250 255 Met Glu Lys Gly Ile Pro Ala Ser Pro Arg Cys Gly Ser Pro ThrAla 260 265 270 Met Glu Ile Asp Lys Arg Gly Ser Pro Thr Pro Gly Thr ArgSer Cys 275 280 285 Leu Ala Pro Ser Leu Gly Leu Phe Gly Ala Ser Leu ThrMet Ala Thr 290 295 300 Glu Val Ala Ala Arg Val Thr Ser Thr Gly Pro HisArg Pro Gln Asp 305 310 315 320 Leu Ala Leu Thr Glu Pro Ser Gly Arg AlaArg Glu Leu Glu Asp Leu 325 330 335 Gln Pro Pro Glu Ala Leu Val Glu ArgGln Gly Gln Phe Leu Gly Ser 340 345 350 Glu Thr Ser Pro Ala Pro Glu ArgGly Gly Pro Arg Asp Gly Glu Pro 355 360 365 Pro Gly Lys Met Gly Lys GlyTyr Leu Pro Cys Gly Met Pro Gly Ser 370 375 380 Gly Glu Pro Glu Val GlyLys Arg Pro Glu Glu Thr Thr Val Ser Val 385 390 395 400 Gln Ser Ala GluSer Ser Asp Ser Leu Ser Trp Ser Arg Leu Pro Arg 405 410 415 Ala Leu AlaSer Val Gly Pro Glu Glu Ala Arg Ser Gly Ala Pro Val 420 425 430 Gly GlyGly Arg Trp Gln Leu Ser Asp Arg Val Glu Gly Gly Ser Pro 435 440 445 ThrLeu Gly Leu Leu Gly Gly Ser Pro Ser Ala Gln Pro Gly Thr Gly 450 455 460Asn Val Glu Ala Gly Ile Pro Ser Gly Arg Met Leu Glu Pro Leu Pro 465 470475 480 Cys Trp Asp Ala Ala Lys Asp Leu Lys Glu Pro Gln Cys Pro Pro Gly485 490 495 Asp Arg Val Gly Val Gln Pro Gly Asn Ser Arg Val Trp Gln GlyThr 500 505 510 Met Glu Lys Ala Gly Leu Ala Trp Thr Arg Gly Thr Gly ValGln Ser 515 520 525 Glu Gly Thr Trp Glu Ser Gln Arg Gln Asp Ser Asp AlaLeu Pro Ser 530 535 540 Pro Glu Leu Leu Pro Gln Asp Pro Asp Lys Pro PheLeu Arg Lys Ala 545 550 555 560 Cys Ser Pro Ser Asn Ile Pro Ala Val IleIle Thr Asp Met Gly Thr 565 570 575 Gln Glu Asp Gly Ala Leu Glu Glu ThrGln Gly Ser Pro Arg Gly Asn 580 585 590 Leu Pro Leu Arg Lys Leu Ser SerSer Ser Ala Ser Ser Thr Gly Phe 595 600 605 Ser Ser Ser Tyr Glu Asp SerGlu Glu Asp Ile Ser Ser Asp Pro Glu 610 615 620 Arg Thr Leu Asp Pro AsnSer Ala Phe Leu His Thr Leu Asp Gln Gln 625 630 635 640 Lys Pro Arg ValLys Tyr Arg Thr Ile Trp Lys Val Lys Asn Lys Glu 645 650 655 Arg Glu SerSer Pro Gly Asn Ala Ser Leu Leu Leu Ile Pro Val Thr 660 665 670 Ala AlaThr Gly Ile Arg Val Leu Gly Leu Gly Leu Gly Asp Leu Gly 675 680 685 GluIle Pro Val Tyr Thr Trp Leu Ala Ser Ser Leu Lys Asn Gly Glu 690 695 700Ser Lys Cys Asp Leu Met Glu Trp Tyr Cys Tyr Thr Val Lys His Pro 705 710715 720 Gly Ser Leu Glu Leu His Gly Leu Arg Met Ser Pro Thr Gly Thr Ser725 730 735 Cys Cys Gly Leu Ile Met Ser Ala Pro Lys Gln Glu Leu Asn AlaIle 740 745 750 Glu Leu Ser Tyr Leu Pro Pro Ala Pro Ile Val Val Val ArgLys Ser 755 760 765 Gly Phe Ser Ala Gln Gln Ser Ala Trp Asp Cys Ile LysPro Ser Ser 770 775 780 Pro Ile Arg Asp Arg Val Ala Leu Leu Cys Pro MetGly Phe Lys Ala 785 790 795 800 Lys Gly Leu Tyr Glu Ser Cys Leu Trp HisSer Pro Glu Ser Ser Gly 805 810 815 Ile Arg Gln Lys Gln Cys Cys Ala AlaLeu Ser Trp Ala Leu Lys Gly 820 825 830 Lys Arg Glu Tyr Leu Gln Gln TyrSer Gly Trp Met Trp Val Pro Gly 835 840 845 Leu Leu Ile Leu Gly Leu GlyLeu Ser Glu Ile His Arg Ser Ser Leu 850 855 860 Gln Val Gln Pro Ala GlyGly Val His Thr Glu Ala Ala Ala Pro Gly 865 870 875 880 Ala Pro Gly HisGln Gly Ala Met Ser Val Thr Tyr Asp Ala Leu Arg 885 890 895 Glu Lys GlnGln Leu Ser Lys Val Gly Asp Leu Pro Ala Leu Thr Trp 900 905 910 Pro GlyPro Leu Ile Ser Gln Met Pro Gly Val Leu Asp Ser Cys Arg 915 920 925 LeuCys Ser Leu Gly Asp Ile Glu Lys Ser Lys Ser Trp Arg Lys Ile 930 935 940Lys Asn Met Val His Trp Ser Pro Phe Val Met Ser Phe Lys Lys Lys 945 950955 960 Tyr Pro Trp Ile Gln Leu Ala Gly His Ala Gly Ser Phe Lys Ala Ala965 970 975 Ala Asn Gly Arg Ile Leu Lys Lys His Cys Glu Ser Glu Gln ArgCys 980 985 990 Leu Asp Arg Leu Met Val Asp Val Leu Arg Pro Phe Val ProAla Tyr 995 1000 1005 His Gly Asp Val Val Lys Asp Gly Glu Arg Tyr AsnGln Met Asp Asp 1010 1015 1020 Leu Leu Ala Asp Phe Asp Ser Pro Cys ValMet Asp Cys Lys Met Gly 1025 1030 1035 1040 Ile Arg Gln Gln Gln Asp PheAla Gly Asp His Met Glu Asn Asn Pro 1045 1050 1055 Ser Gly Val His SerAsp Leu Ala Lys Lys Ala Gly Glu Cys Gly Glu 1060 1065 1070 Gly Leu SerLeu Thr Phe Leu Trp Ala Ser Arg Pro Thr Ile Gln Leu 1075 1080 1085 AlaPro Pro Val Asp Ile Ser Pro Gln Pro Leu Ser Ser Pro Gly Gln 1090 10951100 Thr Tyr Leu Glu Glu Glu Leu Thr Lys Ala Arg Lys Lys Pro Ser Leu1105 1110 1115 1120 Arg Lys Asp Met Tyr Gln Lys Met Ile Glu Val Asp ProGlu Ala Pro 1125 1130 1135 Thr Glu Glu Glu Lys Ala Gln Arg Ala Val ThrLys Pro Arg Tyr Met 1140 1145 1150 Gln Trp Arg Glu Thr Ile Ser Ser ThrAla Thr Leu Gly Phe Arg Ile 1155 1160 1165 Glu Gly Ile Lys Leu Arg GlySer Ala Trp Gly Ala Leu Pro Thr Ala 1170 1175 1180 Pro Gly Ser Arg ProLeu Leu His Pro Gly Leu Leu Pro Gln Pro Gln 1185 1190 1195 1200 Val LeuPro Val Leu Ser Lys Ala Ala Thr Lys Glu Asp Gly Thr Val 1205 1210 1215Asn Arg Asp Phe Lys Lys Thr Lys Thr Arg Glu Gln Val Thr Glu Ala 12201225 1230 Phe Arg Glu Phe Thr Lys Gly Asn His Asn Ile Leu Ile Ala TyrArg 1235 1240 1245 Asp Arg Leu Lys Ala Ile Arg Thr Thr Leu Glu Val SerPro Phe Phe 1250 1255 1260 Lys Cys His Glu Val Ile Gly Ser Ser Leu LeuPhe Ile His Asp Lys 1265 1270 1275 1280 Lys Glu Gln Ala Lys Val Trp MetIle Asp Phe Gly Lys Thr Thr Pro 1285 1290 1295 Leu Pro Glu Gly Gln ThrLeu Gln His Asp Val Pro Trp Gln Glu Gly 1300 1305 1310 Asn Arg Glu AspGly Tyr Leu Ser Gly Leu Asn Asn Leu Val Asp Ile 1315 1320 1325 Leu ThrGlu Met Ser Gln Asp Ala Pro Leu Ala 1330 1335

We claim:
 1. An isolated nucleic acid molecule comprising (a) a nucleicacid molecule comprising a nucleic acid sequence that encodes an aminoacid sequence of SEQ ID NO: 160 through 282; (b) a nucleic acid moleculecomprising a nucleic acid sequence of SEQ ID NO: 1 through 159; (c) anucleic acid molecule that selectively hybridizes to the nucleic acidmolecule of (a) or (b); or (d) a nucleic acid molecule having at least60% sequence identity to the nucleic acid molecule of (a) or (b).
 2. Thenucleic acid molecule according to claim 1, wherein the nucleic acidmolecule is a cDNA.
 3. The nucleic acid molecule according to claim 1,wherein the nucleic acid molecule is genomic DNA.
 4. The nucleic acidmolecule according to claim 1, wherein the nucleic acid molecule is amammalian nucleic acid molecule.
 5. The nucleic acid molecule accordingto claim 4, wherein the nucleic acid molecule is a human nucleic acidmolecule.
 6. A method for determining the presence of a breast specificnucleic acid (BSNA) in a sample, comprising the steps of: (a) contactingthe sample with the nucleic acid molecule according to claim 1 underconditions in which the nucleic acid molecule will selectively hybridizeto a breast specific nucleic acid; and (b) detecting hybridization ofthe nucleic acid molecule to a BSNA in the sample, wherein the detectionof the hybridization indicates the presence of a BSNA in the sample. 7.A vector comprising the nucleic acid molecule of claim
 1. 8. A host cellcomprising the vector according to claim
 7. 9. A method for producing apolypeptide encoded by the nucleic acid molecule according to claim 1,comprising the steps of (a) providing a host cell comprising the nucleicacid molecule operably linked to one or more expression controlsequences, and (b) incubating the host cell under conditions in whichthe polypeptide is produced.
 10. A polypeptide encoded by the nucleicacid molecule according to claim
 1. 11. An isolated polypeptide selectedfrom the group consisting of: (a) a polypeptide comprising an amino acidsequence with at least 60% sequence identity to of SEQ ID NO: 160through 282; or (b) a polypeptide comprising an amino acid sequenceencoded by a nucleic acid molecule comprising a nucleic acid sequence ofSEQ ID NO: 1 through
 159. 12. An antibody or fragment thereof thatspecifically binds to the polypeptide according to claim
 11. 13. Amethod for determining the presence of a breast specific protein in asample, comprising the steps of: (a) contacting the sample with theantibody according to claim 12 under conditions in which the antibodywill selectively bind to the breast specific protein; and (b) detectingbinding of the antibody to a breast specific protein in the sample,wherein the detection of binding indicates the presence of a breastspecific protein in the sample.
 14. A method for diagnosing andmonitoring the presence and metastases of breast cancer in a patient,comprising the steps of: (a) determining an amount of the nucleic acidmolecule of claim 1 or a polypeptide of claim 6 in a sample of apatient; and (b) comparing the amount of the determined nucleic acidmolecule or the polypeptide in the sample of the patient to the amountof the breast specific marker in a normal control; wherein a differencein the amount of the nucleic acid molecule or the polypeptide in thesample compared to the amount of the nucleic acid molecule or thepolypeptide in the normal control is associated with the presence ofbreast cancer.
 15. A kit for detecting a risk of cancer or presence ofcancer in a patient, said kit comprising a means for determining thepresence the nucleic acid molecule of claim 1 or a polypeptide of claim6 in a sample of a patient.
 16. A method of treating a patient withbreast cancer, comprising the step of administering a compositionaccording to claim 12 to a patient in need thereof, wherein saidadministration induces an immune response against the breast cancer cellexpressing the nucleic acid molecule or polypeptide.
 17. A vaccinecomprising the polypeptide or the nucleic acid encoding the polypeptideof claim 11.